Vous êtes sur la page 1sur 10

Effects of Different

Dietary Proteins and


Amino Acids on Skeletal
Muscle Hypertrophy in
Young Adults After
Resistance Exercise: A
Systematic Review
Henning Langer, BA and Anja Carlsohn, PhD
Universita t Potsdam, Professur Sportmedizin und Sportorthopa die, Potsdam, Germany
A B S T R A C T
THIS ARTICLE REVIEWS THE
AVAILABLE LITERATURE ON
WHICH PROTEINS, AMINO ACIDS,
OR COMBINATION OF BOTH
SEEM TO BE OPTIMAL TO
ENHANCE HYPERTROPHY AFTER
RESISTANCE EXERCISE IN
YOUNG ADULTS. DEPENDING ON
THE CONTENT OF ESSENTIAL
AMINO ACIDS AND PARTICU-
LARLY LEUCINE, EITHER AN
IMMEDIATE INGESTION OF ;20 G
MILK PROTEIN FOLLOWED BY A
SIMILAR AMOUNT ;1 HOUR
LATER, OR A SINGLE BOLUS
OF ;40 G SEEMS TO BE SUIT-
ABLE. GREATER AMOUNTS
MIGHT BE NECESSARY IF A
PROTEIN OF LOWER QUALITY IS
CHOSEN (I.E., PLANT-BASED
PROTEINS) TO MATCH THE
REQUIRED AMINO ACID QUANTI-
TIES AND FACILITATE MUSCLE
GROWTH.
INTRODUCTION
I
nvestigating skeletal muscle hyper-
trophy is important for numerous
reasons. Recreational and profes-
sional athletes can benet from hypertro-
phy as a muscle with increased cross-
sectional area (CSA) can exert more
force, eventually leading to greater
strength and power potential (39). Muscle
hypertrophy has vital implications not
only for performance but also from
a health perspective. Muscle tissue plays
a major role in regulating our metabolism
and consequentially all diseases that are
related to it (34,43). Furthermore, building
up muscle tissue in young subjects could
be a promising intervention to battle
increasingly prevalent diseases such as
sarcopenia or cachexia, which are related
to muscle loss and muscle weakness
(5,52). Starting early enough with this
could be particularly important, as
humans have been reported to become
resistant to anabolic stimuli during aging
(10,33,36,65). How to optimally stimulate
skeletal muscle hypertrophy in young
healthy adults through exercise is a con-
troversial topic (14).
Skeletal muscle hypertrophy occurs
when the aggregate of muscle protein
synthesis (MPS) exceeds the aggregate
of muscle protein breakdown (MPB)
for an extended period of time, resulting
in a positive net protein balance (NPB).
It is well-established that in order to
stimulate this process, it is essential to
overload the muscle (27,28). However,
it has been shown that in a fasted state
and without sufcient protein intake,
the anabolic effects after RE are dimin-
ished, as both MPS and MPB rise in
a similar fashion resulting in a negative
NPB (1,22,46,60).
Essential amino acids (EAAs) play a key
role, as they increase MPS and minimize
MPB, causing a higher NPB compared
with a mixture of nonessential amino
KEY WORDS:
milk; whey; casein; soy; leucine; pro-
tein; resistance training; hypertrophy
Copyright National Strength and Conditioning Association Strength and Conditioning Journal | www.nsca-scj.com
33
acids (AAs) (60). The EAA leucine has
been suggested as a MPS key modulator
by increasing the activity of important
signaling proteins (19). Sufcient protein
intake seems to be important to maxi-
mize the contractile protein accretion
after RE, and this seems to be partially
dependent on the EAA content of the
consumed protein. A protein high in
EAA and with a high digestible indis-
pensable amino acid score may be
termed as a high-quality protein (23).
Examples of high-quality protein sources
include whey-, soy-, casein-, multicom-
ponent proteins or even AA supple-
ments such as branched-chain amino
acids (BCAA) or pure leucine. This
review will overview the effects of differ-
ent dietary proteins ingested immedi-
ately before, during, or after RE. It has
been shown that the anabolic effects of
RE differ depending on certain factors,
such as level of experience of the sub-
jects, training volume, and training inten-
sity. Therefore, the protocols of the
reviewed studies will be elucidated in
detail to allow for the results to be put
in an appropriate perspective.
SEARCH STRATEGY
A systematic PubMed database search
was performed using combinations of
the following terms: protein ingestion,
casein, whey, soy, egg, beef,
leucine, amino acids, resistance train-
ing, resistance exercise, and weight-
lifting. Language of the studies was
limited to English (Figure).
CRITERIA
This review included studies that
looked at the effects of more than
one protein or AA source during their
respective experiments. The subjects in
those studies had to undergo at least 1
bout of intense RE, and indicators of
skeletal muscle hypertrophy had to be
measured before and after the inter-
vention. Examples of such indicators
are acute changes in muscle protein
turnover or long-term changes in body
composition and muscle size. All stud-
ies administered the protein in a time
frame of 90 minutes before, during, or
after RE as this seems to be crucial for
the hypertrophic response (1,22,45,53).
Studies with older (+40 years) or
diseased subjects were excluded
(Table 1).
A total of 5,868 articles were initially
found. Of this, 12 studies till July 2013
fullled the dened inclusion criteria
and were included in this review.
DIFFERENT PROTEIN SOURCES
ON MUSCLE PROTEIN TURNOVER
AND HYPERTROPHY
Seven studies analyzed milk proteins
(whey and casein) and their different
effects on postexercise muscle anabo-
lism or chronic changes in body com-
position. Four of them compared the
results with soy protein.
Tipton et al. (59) included 23 healthy,
young, untrained subjects of both gen-
ders. Subjects had to perform a single
bout of leg extension, consisting of 10
sets of 8 repetitions (reps) at 80% of
the 1 repetition maximum (1RM) of
the subjects, with a 2-minute break
between each set. One hour after this
bout, 20 g of whey protein (2.3 g leucine),
casein protein (1.7 g leucine), or placebo
(water) were ingested by the subjects.
Vastus lateralis biopsies were taken
55 minutes before, immediately before,
and 120 minutes and 300 minutes after
RE. Leg blood ow was measured
4535 minutes before RE and at
4045 minutes, 8090 minutes, 115
120 minutes, 200210 minutes, and
290300 minutes after exercise. Blood
samples were taken at 17 points from
35 minutes before exercise to 300 mi-
nutes after exercise. Appearance of AAs
(phenylalanine and leucine) in the blood
and muscle was measured, and NPB
was calculated using the following for-
mula: NPB 5 (blood ow across the
leg) 3 (arterial AA concentration 2
venous AA concentration).
Whey protein ingestion resulted in
a more rapid increase and larger peak
in leucine concentrations in blood and
muscle than casein ingestion, likely
because of different digestive properties
of the proteins (2,20,21). However, in
terms of the potential hypertrophy
(NPB), no signicant difference between
the groups was observed. The placebo
had no effect on the values.
Wilkinson et al. (64) compared a milk
protein containing drink (fat-free milk)
with an isonitrogenous and isoenergetic
soy protein drink. Eight young subjects
who were regularly engaged in RE ($4
days per week) participated in 2 bouts of
a unilateral leg workout, separated by $1
week. Three standardized exercises (leg
press, leg curl, and leg extension) were
executed, each for 4 sets at 80% of the
1RM with 2-minute rest between each
set. After the exercise, either the soy or
milk drink was ingested. Both included
18.2 g of protein, 1.5 g of fat, and 23 g of
carbohydrates (CHO). Muscle biopsies
were taken immediately before and after
RE, at 60 minutes, 120 minutes, and
180 minutes after workout. Blood sam-
ples were drawn 90 minutes before exer-
cise, directly before and after exercise,
and 30, 60, 90, 120, and 180 minutes after
RE. Blood ow was measured before
and after workout, as well as every hour
after RE up to 180 minutes. Similarly to
Tipton et al. (59), fractional synthesis rate
(FSR in percentage per hour) was calcu-
lated as described by Phillips et al. (46).
NPB was calculated by subtracting frac-
tional breakdown rate from FSR. The
fat-free milk stimulated FSRto a substan-
tially greater extent (34%) than the soy
drink, resulting in a signicantly greater
NPB area under the curve (AUC). This
indicates that milk proteins are a better
stimulator of postexercise muscle anab-
olism in young resistance athletes than
soy protein.
In a follow-up study by Hartman et al.,
56 college students who were previ-
ously not experienced in RE completed
a 12-week trial. Before the start of the
program, subjects were randomly as-
signed to 1 of the 3 groups (milk, soy,
or control) (29). Each subject was
trained 5 days per week on a rotating
split program. The split program con-
sisted of 3 different types of sessions,
for a total of 60 RE sessions over the
course of the 12 weeks. During the trial,
load was progressively increased (start-
ing at 80%1RM), whereas the number of
reps per set decreased. Immediately after
each workout and again 1 hour later,
each subject consumed a drink. The
drink was either skim milk (17.5 g of
Effects of Different Dietary Proteins
VOLUME 36 | NUMBER 3 | JUNE 2014
34
protein, 25.7 g of CHO and 0.4 g of fat),
an isoenergetic and isonitrogenous soy
beverage, or an isoenergetic control
drink consisting of maltodextrin.
Changes in fat- and bone-free mass
and type 1 and type 2 ber CSA were
measured using dual-energy x-ray ab-
sorptiometry (DEXA) and muscle biop-
sies of the vastus lateralis, respectively.
Both methods were used twice: before
the start of the RE regimen and after the
completion of the 12-week program.
The greatest gains in lean mass and
CSA could be observed in the milk pro-
tein group, followed by the soy protein
and the control groups. These ndings
suggest that a combination of milk pro-
teins (i.e., whey and casein) is superior to
soy protein or CHO alone in terms of
promoting hypertrophy in young resis-
tance trained men.
Based on those ndings, Candow et al.
(13) hypothesized that whey protein
would support fat-free mass gains in
young adults to a greater degree than
soy protein or an isoenergetic maltodex-
trin supplement. Twenty-seven untrained
subjects were randomly assigned into 3
groups: whey protein, soy protein, and
placebo (maltodextrin). Each participant
received a prepacked bag containing one
of the supplements. They were instructed
to ingest 1.2 g per kg bodyweight of the
supplement, 0.4 g per kg bodyweight
before and after RE, as well as before
bed. The weight training program was
composed of a 3-day rotating split, with
69 exercises per session and 45 sets per
exercise at 6090%of their 1RMfor 612
reps, for a total study duration of 6 weeks.
All participants had to report their die-
tary intake of 3 days during the rst and
last weeks. No signicant difference in
dietary habits between the groups or
weeks was observed. Mean calorie in-
takes ranged from 2,978 6 702 to
3,129 6 591 kcal per day and protein
intakes from 1.6 6 1.3 to 1.9 6 1.3 g
per kg bodyweight per day. However,
it is important to mention that the sup-
plementary protein intake of the whey
and soy groups was not included in these
data, meaning that most subjects were
likely to not only match but also clearly
exceed the recommended protein re-
quirements for athletes engaged in RE
(37,38,47,50). As a result of this trial,
the whey and soy protein groups showed
signicantly greater gains in LBM (mea-
sured by DEXA) than the placebo group.
In this study, the hypothesis that whey
Table 1
Inclusion and exclusion criteria
Criteria Included Excluded
Age ,40 y .40 y
Protein/amino acid source .1 #1
Health status Healthy Diseased
Timed ingestion 690 min of RE .690 min of RE
Resistance exercise $1 bout per study No RE
Indicators of muscle
growth measured
Yes No
Figure. Flow chart of the process of identifying and including studies for the systematic literature review.
Strength and Conditioning Journal | www.nsca-scj.com
35
protein would support muscle growth
to a greater extent than soy protein
could not be conrmed.
Tang et al. (56) investigated whether
there is a different effect for the isolated
proteins of milk (whey and casein) com-
pared with soy. In contrast to Tipton
et al. (59), this study found a signicant
difference between casein, soy, and
whey on MPS. Eighteen young and
healthy subjects experienced in weight-
lifting (23 sessions per week) com-
pleted this trial. A single bout of 2
unilateral leg exercises was performed:
4 sets of leg press and leg extension at
100% of their previously determined
1012 RM with 2-minute rest between
each set. Immediately after RE, a drink
of 21.4 g of whey, 21.9 g of casein, or
22.2 g of soy protein was ingested. Each
drink contained approximately 10 g of
EAA. A continuous L-[ring-
13
C
6
] phe-
nylalanine infusion, blood samples, and
muscle biopsies were used to measure
MPS at rest, after the consumption of
the drink and up to 180 minutes after
the workout. Despite similar EAA con-
tent in all drinks, the ingestion of the
whey drink resulted in a signicantly
greater AUC of leucine and higher
MPS than the ingestion of soy or casein.
Therefore, whey protein seems to be
a more potent stimulator of acute mus-
cle anabolism after RE than casein or
soy. Several authors suggested that this
might be because of the faster absorp-
tion rate of whey when compared with
other proteins that are similar in EAA
content (2,20,21). However, an infusion
protocol of only 3 hours postprandially
is unlikely to catch the slow acting
effects of casein on MPS and could
therefore show an incomplete picture.
Reitelseder et al. (49) conducted
another study comparing whey with
casein. They conrmed what Tipton
et al. (59) found and could not replicate
Tang et al. (56). They had 17 subjects
not experienced in RE perform 10 sets
of 8 reps at 80% of their 1RM with 3-
minute rest between the sets. After RE,
a drink of either whey, casein (0.3 g per
kilogram lean bodyweight), or control
(non caloric) was ingested. MPS was
measured in a similar way as described
in earlier studies (46,56,59,64). Com-
pared to the study by Tang et al., the
infusion protocol of this investigation
was twice as long and assessed changes
in MPS over a 6 hour period. Whey
ingestion was followed by a greater
peak in MPS during the early phase
(13 hours), whereas casein caused
a higher elevation during the later phase
(36 hours), resulting in a similar mean
MPS over the total period (16 hours).
This is supporting the idea that the
slower absorption kinetics of casein re-
quires a longer infusion period to allow
for a comprehensive comparison to its
effects on MPS versus whey protein.
However, it was a special form of casein
that was used in this trial. Unlike micel-
lar casein that is found in bovine milk
and was used in the other experiments
mentioned above, this form (calcium
caseinate) has different digestive prop-
erties that are more similar to whey pro-
tein. Micellar casein is not acid soluble
causing it to be digested more slowly,
whereas caseinates are acid soluble and
release their AAs much more rapidly
into the intestine, bloodstream, and ulti-
mately peripheral tissues (18). There-
fore, the results of this study cannot
be applied to the most common form
of casein (micellar casein) and are likely
to be directly linked to this particular
form of casein (caseinate).
Recently, another comparison between
whey and casein did not show any sig-
nicant differences between changes in
body composition (63). Wilborn et al.
had 16 female basketball players partic-
ipate in this trial of 8 weeks. They par-
ticipated in 4 whole-body RE sessions
per week, which were not specied in
terms of intensity. Subjects performed 3
conditioning units per week suited
toward basketball. The subjects were
randomized into a whey or casein pro-
tein group. Both groups consumed
either an isocaloric portion of 24 g
casein or whey immediately after each
of the 7 training sessions of the week. By
the end of the 8 weeks, both groups had
increased muscle mass and strength in
a similar manner. However, the lack of
a nonprotein control group makes it
difcult to distinguish the effects of
the training protocol from the protein
supplementation.
Taken together, these results indicate
that a combination of milk proteins
as it is naturally occurring in bovine
milk seems to be superior to soy pro-
tein in promoting hypertrophic gains
in young healthy adults (29,56,64).
When looking at the effects of the iso-
lated milk proteins, whey protein
seems to be more facilitating in the
early phase after RE, whereas casein
ingestion results in a slower, but more
prolonged effect on MPS (49,56).
It has been suggested that the main rea-
son for soy protein being less potent in
augmenting MPS, despite its high-quality
AA prole and fast digestion rate, is the
way it is partitioned. Its AAs seem to be
primarily distributed to the splanchnic
region and less toward peripheral tissues
like muscle (3,24,57).
THE ROLE OF LEUCINE AND
OTHER AMINO ACIDS
Results from Wilkinson and Hartman
et al. suggest that milk may be an ef-
cient postworkout nutrition and protein
source (29,64). The protein in bovine
milk as used in those studies usually
contains 20% whey to 80% casein pro-
tein (31). A combination of whey and
casein protein could be a promising
workout beverage. However, substantial
differences in absorption kinetics and
time effects of whey and casein were
observed, leaving the question whether
it is possible to ameliorate the effects of
fast- and slow-acting proteins on lean
mass accretion by combining them.
The exact role of AAs within proteins
or added to them remained unclear.
Kerksick et al. (35) compared the
effects of 3 different postworkout
drinks on body composition after 10
weeks of RE. Thirty-six subjects were
assigned to either a whey and casein
protein blend (48 g: 40 g of whey and
8 g of casein), a whey protein that was
stacked with BCAAs and glutamine
(48 g: 40 g of whey, 3 g of BCAAs +
5 g of L-glutamine), or an isoenergetic
CHO placebo that had to be ingested
in a 2-hour window after RE or in the
morning (9 AM) on training-free days.
Effects of Different Dietary Proteins
VOLUME 36 | NUMBER 3 | JUNE 2014
36
The participants were experienced in
weightlifting and completed a 10-week
RE program (4 sessions per week), with
78 exercises and 2124 sets per session,
respectively. Over the course of the
study, the training loads were progres-
sively increased, and each set throughout
the entire program was executed until
the subjects could not perform another
complete repetition. The preplanned
reps ranged from 6 to 10 per set for
most of the exercises and 25 per set
for abdominal exercises. Similarly to
Candow et al. (13), the subjects were
instructed to report their dietary habits
over a 4-day period before the start of
the trial, during weeks 2, 5, 8, and 10 of
the program. Energy intake from group
to group and week 010 varied between
29.2 and 39.8 kcal per kilogram body-
weight and protein intake from 1.4 to
2.5 g per kilogram bodyweight, respec-
tively. Although there was no statistical
difference between the groups, it is
important to note that every single
subject was close to matching or
even exceeding the recommended pro-
tein intake per day for RE athletes
(37,38,47,50). As a result of this trial,
signicant gains in LBM measured by
DEXA occurred in the whey plus casein
group only, indicating that this protein
blend would be advantageous compared
with whey protein that was stacked with
EAA and glutamine, which had no
effect on body composition.
In an attempt to further elaborate on
the role of leucine and EAA on MPS,
Churchward-Venne et al. (16) looked at
the effects of different whey protein dos-
ages, with or without added leucine or
EAAs. A regular dose of whey protein
(25 g) was compared with 2 different
groups of suboptimal dosages of whey
protein (6.25 g). One group had added
leucine, to match the leucine content of
the regular dosage in it and the other
had added EAAs in it, to match every
EAA of the regular dosage besides
leucine. They hypothesized that adding
leucine, but not EAA, would result in
a MPS response similar to the group
with the complete whey (19,51,60).
They had 24 subjects not experienced
in RE performa single bout of unilateral
exercise, consisting of 4 sets of 1012
reps leg extension and leg press at 95%
of their previously determined 1RM.
Immediately after this bout, they were
assigned to 1 of the 3 groups in a single-
blinded fashion and ingested the bever-
age. MPS was measured as described
earlier, using infusion, blood samples,
and a biopsy from the worked muscle
(vastus lateralis) (46,49,56,59,64). As
a result, they observed equal MPS
responses between the groups in the
early hours (13 hours) after RE, but
only the complete whey group (25 g)
was able to sustain this response for
a prolonged time (35 hours after exer-
cise). This implies that despite the fact
that leucine was suggested to be indis-
pensable for MPS and NPB to occur,
less of it than previously thought might
be necessary when other EAAs are
available in sufcient amounts. In addi-
tion, whey in adequate quantities seems
to be a better choice postworkout than
little amounts of protein stacked with
leucine or other EAAs. Whey in ade-
quate quantities seems to be a better
choice after workout than little
amounts of protein stacked with leucine
or other EAAs.
Reidy et al. (48) compared the effects
of whey protein with a protein blend
consisting of sodium caseinate (50%),
whey, and soy (25% each). Both bev-
erages contained approximately the
same amount of leucine (;1.8 g) and
EAAs (;8.7 g). Nineteen untrained
subjects completed a single bout of
leg exercise. They performed 8 sets of
10 reps leg extension at increasing
intensity from 55 to 70% of their
1RM with 3-minute rest between
the sets. FSR was measured using
similar methods as described in earlier
studies (16,46,49,56,59,64). The protein
drink was ingested 1 hour after RE.
Depending on the subjects group
and their bodyweight (0.30.35 g per
kilogram bodyweight), they either
received ;18 g of whey or ;19 g of
the protein blend. The group hypoth-
esized that the individual characteris-
tics of each component of the protein
blend would yield unique advantages
over whey protein. No signicant
difference in MPS between the groups
could be observed in the early phase
after RE. However, as expected, the
slower kinetics of the caseinate and
the plant protein led to an increased
MPS during the late phase (24 hours
after RE), whereas no increase in MPS
could be observed for whey-only at that
time. This shifted effect of the slower
proteins is also reected by differences
in the rise and fall of AA concentrations
in the serum. The authors therefore sug-
gested that a multicomponent protein
could be a better choice after RE. Sim-
ilar to the study by Reitelseder et al. (49),
it is important to consider that sodium
caseinate just like calcium caseinate does
not resemble the digestive characteris-
tics of micellar casein and instead is
absorbed much faster (18).
Herda et al. compared bioenhanced
whey (20 g of whey + 5 g of leucine)
with normal whey (20 g), a placebo (27
g of maltodextrin), and a control group
(30). Over 8 weeks of RE, 106 subjects,
49 of them experienced in weight train-
ing, trained 3 times per week at 80% of
their 1RM with 2-minute rest between
the sets. The subjects were randomly
assigned into 5 different groups: bioen-
hanced whey combined with a low-
volume RE program, bioenhanced
whey plus moderate volume, normal
whey plus moderate volume, placebo
plus moderate volume, and control
plus moderate volume. The different
beverages were ingested 30 minutes
before and after workout. On rest days,
they were instructed to drink it in the
morning in a postabsorptive state. Each
participant had to report 3 random days
of dietary intake (2 week days and 1
weekend day) and was asked to main-
tain his habits throughout the study.
Average kilocalorie intake between the
groups ranged from 2,320 6129 kcal to
3,029 6 181 kcal. Protein intake varied
from between 0.691.96 g per kilogram
bodyweight (lowest average group
intake) to 1.262.86 g per kilogram
bodyweight (highest average group
intake), respectively. The placebo and
control groups consumed signicantly
less protein than the other 3 groups.
However, even in those groups some
Strength and Conditioning Journal | www.nsca-scj.com
37
individuals excessively exceeded the
recommended (37,38,47,50) protein
intake per day. The changes in body
composition were evaluated using
peripheral quantitative computed tomog-
raphy. No signicant changes in muscle
CSA or LBM gains could be observed
between the groups, despite the differ-
ence in total protein intake.
Recently, Joy et al. (32) compared the
effects of 48 g of rice protein versus 48
g of whey protein after RE for 8 weeks.
Subjects were 24 healthy young
males who were experienced in strength
training for at least 1 year. The RE
protocol consisted of an undulating peri-
odization program for 3 days a week.
They performed alternating whole-
body sessions at either their 812 RM
or 25 RM. The weights were increased
by 25% when the desired number of
reps was achieved. The authors hypoth-
esized that there would be no difference
between the groups, agreeing with
earlier assumptions about a leucine
threshold (4). Indeed, no signicant dif-
ference between the groups was found.
Table 2
Protein sources, duration, and outcome
Reference Protein and amino acid content
of the beverages
Indicators of hypertrophy Duration Results
Tipton et al. (59) 20 g whey or 20 g casein NPB Single bout NPB[; no difference between
the groups
Wilkinson et al. (64) 18.2 g milk protein or 18.2 g soy
protein
NPB Single bout NPB[; milk . soy
Hartman et al. (29) 17.5 milk protein or 17.5 soy
protein
CSA, LBM 12 wk CSA[, LBM[, milk . soy
Candow et al. (13) ;30 g whey, ;30 g soy protein
or ;30 g maltodextrin
LBM 40 d LBM[; no difference between
the groups
Tang et al. (56) 21.4 g whey, 21.9 g casein, or
22.2 g soy protein
MPS Single bout MPS whey . soy . casein
Reitelseder et al.
(49)
;20 g whey or ;20 g casein MPS Single bout MPS no difference between
the groups
Wilborn et al. (63) 24 g whey or 24 g casein LBM 8 wk LBM[; no difference between
the groups
Kerksick et al. (35) 40 g whey + 8 g casein or 40 g
whey + 3 g BCAAs + 5 g
glutamine
LBM 10 wk LBM[; whey plus casein .
whey plus BCAA and
glutamine
Churchward-Venne
et al. (16)
25 g whey, 6.25 g whey + leucine
(matched with 25 g whey),
6.25 g whey + EAA (matched
with 25g whey, besides
leucine)
MPS Single bout MPS[; no difference between
the groups in the early
response; MPS remained
elevated during the late
phase only in the 25 g whey
group
Reidy et al. (48) ;18 g whey or ;19 g protein
blend (25% whey, 50%
sodium caseinate, 25% soy)
MPS Single bout MPS[; no differences between
the groups in the early
response; MPS remained
elevated during the late
phase only in
multicomponent protein
group
Herda et al. (30) 20 g whey, 20g + 5 g leucine,
27 g maltodextrin or placebo
CSA, LBM 8 wk LBM[; no difference between
the groups
Joy et al. (32) 48 g whey or 48 g rice protein LBM 8 wk LBM[; no difference between
the groups
Effects of Different Dietary Proteins
VOLUME 36 | NUMBER 3 | JUNE 2014
38
The rice group increased their lean mass
by 2.5 kg, which was measured using
DEXA. The whey group gained 3.2 kg
of lean mass during the intervention.
Both increased their 1RM of the per-
formed exercises in a similar manner.
The authors concluded that the study
provides evidence for the theory of
a leucine threshold. They assume that
this threshold is within the range
of 1.73.5 g of leucine. Similar to
Wilborn et al., this study was limited
by the lack of a control group.
DISCUSSION
Although in contrast to the majority
of the ndings, the results of Herda
et al. are in line with a number of
other studies that could not nd any
positive effects for protein or AA sup-
plementation combined with RE
(7,12,25,26,61,62). However, it is the
only study fullling the criteria of this
review that failed to show a positive
effect of protein supplementation on
indicators of skeletal muscle hyper-
trophy. There are two main reasons
that could explain these results. One
is that most subjects in the study of
Herda et al. were not experienced in
RE, and it is a common observation
that novice lifters tend to respond
markedly well to the onset of RE,
unlike advanced athletes. This might
be because of a decreased MPS sig-
naling in advanced athletes, caused
by chronic loading of the muscles
(17,41,42,54). It is possible that the
beginners in the study by Herda
et al. could have beneted from their
initial sensitivity to RE, resulting in
signicant hypertrophy despite sub-
optimal nutritional circumstances.
The authors agree on the chance that
the naturally high basal MPS rates in
young men, in conjunction with an
efcient RE program, could have
already been such a great stimulus
that all additional effects of protein
supplementation got whitewashed
(30). Additionally, some of the sub-
jects in each of the groups exceeded
the current recommendations for
daily protein intake by a great margin.
Potentially, this could have slightly
distorted the results despite a signi-
cant mean difference in overall pro-
tein intake between the groups.
Yet it is possible that overall daily protein
and calorie intake are paramount to pro-
tein ingestion with training. However,
there is no doubt that a high-quality pro-
tein source enhances acute RE-induced
MPS and NPB (16,48,49,56,59,64).
Moreover, acute NPB after exercise
and EAA ingestion reects 24-hour
NPB (58), and there is evidence for pro-
tein to augment long-term LBM gains to
a greater degree than placebo or CHO:
A recent meta-analysis has shown that
improvements in LBM favor protein
supplementation over placebo (15). This
was regardless of the age or training sta-
tus of the subjects (15). This indicates
that studies unable to nd an effect for
protein supplementation to further
enhance the effects of REmay have been
underpowered.
Eventually, based on the current liter-
ature, isolated or mixed milk proteins
and regular bovine milk seem to have
the most potential to facilitate muscle
growth compared with other proteins
(29,49,56,64).
Bovine milk is, despite its high casein
content, rapidly absorbed, resulting in
an acute MPS response that is surpris-
ingly close to isolated whey (64). The
growth-facilitating effects of milk pro-
teins seem to be greater than for other
proteins not only on the short-term but
also on the long-term in young healthy
adults (29,56,64). Furthermore, both
components of milk protein (whey
and casein) are high in leucine and
other EAAs, but only whey seems to
effectively augment acute postworkout
MPS because of its fast absorbable
characteristics (56).
In general, it appears that combining
both milk proteins, as it naturally hap-
pens in bovine milk, could be a promis-
ing strategy in further optimizing the
MPS response after RE. The fast-
acting characteristics of whey are com-
plemented by the slow-acting charac-
teristics of casein, resulting in a rapid,
but also prolonged increase in MPS
(49,64). This seems to be the case
regardless of the type of casein
(29,35,64). Adding other proteins such
as soy to those milk proteinbased
blends might be advantageous as well,
but further research is needed to con-
rm the benets of such multicompo-
nent proteins (48).
Moreover, adding leucine or other
AAs to a sufcient amount of protein,
which is already high in EAA, does
not seem to further augment skeletal
muscle hypertrophy (30,35). However,
if the amount of high-quality protein
and leucine is below what seems to be
the sufcient amount (20 g and 11.7 g,
respectively) (4,32,40), adding EAA or
leucine could help to still get the opti-
mal, early response after RE (16). Alter-
natively, exceeding those amounts and
consuming enough of a protein that is
poor in leucine until one transgresses
the proposed threshold of 11.7 g could
be feasible as well, if a protein of higher
quality is not available (32). However, if
the amount of protein is insufcient and
the shortcoming is compensated with
the addition of leucine or EAA, it would
be advisable to endorse such a compro-
mise with a full dose of a high-quality
protein shortly afterward, as only a com-
plete protein seems to allow for a pro-
longed MPS (16).
It is difcult to compare acute studies
that measured muscle protein turnover
with long-term interventions which
measured changes in muscle size. Little
is known on the long-term adaptations
of skeletal muscle to different levels of
protein intake and timing. For example,
effect sizes of muscle growth found in
long-term interventions are commonly
substantially smaller than expected
based on acute studies that measured
muscle protein turnover (30,40). Future
experiments should target the physio-
logical changes that might occur
between acute studies and long-term
interventions. For accurate recommen-
dations, further research is needed to
determine the optimal ratio of fast- to
slow-acting proteins and the role of
other AAs besides leucine. Moreover,
other high-quality protein sources such
as beef protein or egg protein have been
shown to ameliorate muscle protein
Strength and Conditioning Journal | www.nsca-scj.com
39
turnover to a similar degree as milk pro-
teins (40,44,55). However, to date, this
was performed exclusively in elderly,
dissociated from RE or without com-
paring it to another protein during the
same trial. Those ndings are yet to be
applied to resistance trained young sub-
jects and longterm interventions.
CONCLUSION
Most of the results suggest that in-
gesting milk protein directly before
or after RE is advantageous for
individuals interested in optimizing
skeletal muscle hypertrophy. The
available literature indicates that
a combination of fast- and slow-
acting milk proteins (i.e., whey and
casein) could provide the most per-
petual anabolic effects. The exact
amount that should be ingested is
dependent on the quality of the pro-
tein and its content of EAA, speci-
cally leucine, with 11.7 g of leucine
appearing to be optimal to facilitate
muscle protein accretion in young
subjects. Approximately 20 g of a milk
protein followed by a similar amount
;1 hour later, or alternatively ;40 g
in a single bolus feeding seem to be
sufcient to maximize the acute MPS
response after RE, as well as long-
term hypertrophy. Exceeding those
amounts might be necessary only if
a protein of lower quality (i.e.,
plant-based protein) is chosen. Par-
ticularly, individuals with low protein
intake and protein sources of poor
quality could benet from protein
supplementation around training.
Conicts of Interest and Source of Funding:
The authors report no conicts of interest
and no source of funding.
Henning Langer
is a graduate
student (MSc
program, Biology
of Human Perfor-
mance and
Health) at Maas-
tricht University
(Netherlands).
Anja Carlsohn is
an Assistant
Professor at the
University of
Education
Schwaebisch
Gmuend
(Germany).
ACKNOWLEDGMENT
The authors thank Fabian Ottawa for
his assistance in revising the article.
REFERENCES
1. Andersen LL, Tufekovic G, Zebis MK,
Crameri RM, Verlaan G, Kjr M, Suetta C,
Magnusson P, and Aagaard P. The effect of
resistance training combined with timed
ingestion of protein on muscle ber size
and muscle strength. Metabolism54: 151
156, 2005.
2. Boirie Y, Dangin M, Gachon P,
Vasson M-P, Maubois J-L, and Beaufre` re B.
Slow and fast dietary proteins differently
modulate postprandial protein accretion.
Proc Natl Acad Sci U S A 94: 14930
14935, 1997.
3. Bos C, Metges CC, Gaudichon C,
Petzke KJ, Pueyo ME, Morens C,
Everwand J, Benamouzig R, and Tome D.
Postprandial kinetics of dietary amino acids
are the main determinant of their
metabolism after soy or milk protein
ingestion in humans. J Nutr 133: 1308
1315, 2003.
4. Breen L and Phillips SM. Skeletal muscle
protein metabolism in the elderly:
Interventions to counteract the anabolic
resistance of ageing. Nutr Metab (Lond)
8: 68, 2011.
5. Breen L and Phillips SM. Interactions
between exercise and nutrition to prevent
muscle waste during ageing. Br J Clin
Pharmacol 75: 708715, 2013.
6. Brown EC, DiSilvestro RA, Babaknia A, and
Devor ST. Soy versus whey protein bars:
Effects on exercise training impact on lean
body mass and antioxidant status. Nutr J 3:
22, 2004.
7. Bucci L, Hickson J Jr, Wolinsky I, and
Pivarnik J. Ornithine supplementation and
insulin release in bodybuilders. Int J Sport
Nutr 2: 287291, 1992.
8. Burd NA, Holwerda AM, Selby KC,
West DW, Staples AW, Cain NE,
Cashaback JG, Potvin JR, Baker SK, and
Phillips SM. Resistance exercise volume
affects myobrillar protein synthesis and
anabolic signalling molecule
phosphorylation in young men. J Physiol
588: 31193130, 2010.
9. Burd NA, Mitchell CJ, Churchward-
Venne TA, and Phillips SM. Bigger weights
may not beget bigger muscles: Evidence
from acute muscle protein synthetic
responses after resistance exercise. Appl
Physiol Nutr Metab 37: 551554, 2012.
10. Burd NA, Wall BT, and van Loon LJ. The
curious case of anabolic resistance: Old
wives tales or new fables? J Appl Physiol
(1985) 112: 12331235, 2012.
11. Burd NA, West DW, Staples AW,
Atherton PJ, Baker JM, Moore DR,
Holwerda AM, Parise G, Rennie MJ, and
Baker SK. Low-load high volume
resistance exercise stimulates muscle
protein synthesis more than high-load low
volume resistance exercise in young men.
PLoS One 5: e12033, 2010.
12. Campbell WW, Crim MC, Young VR,
Joseph LJ, and Evans WJ. Effects of
resistance training and dietary protein
intake on protein metabolismin older adults.
Am J Physiol 268: E1143E1153, 1995.
13. Candow DG, Burke NC, Smith-Palmer T,
and Burke DG. Effect of whey and soy
protein supplementation combined with
resistance training in young adults. Int J Sport
Nutr Exerc Metab 16: 233244, 2006.
14. Carpinelli RN, Otto RM, and Winett RA.
A critical analysis of the ACSM position
stand on resistance training: Insufcient
evidence to support recommended training
protocols. J Exerc Physiol 7: 160, 2004.
15. Cermak NM, Res PT, de Groot LC,
Saris WH, and van Loon LJ. Protein
supplementation augments the adaptive
response of skeletal muscle to resistance-
type exercise training: A meta-analysis. Am
J Clin Nutr 96: 14541464, 2012.
16. ChurchwardVenne TA, Burd NA,
Mitchell CJ, West DW, Philp A,
Marcotte GR, Baker SK, Baar K, and
Phillips SM. Supplementation of
a suboptimal protein dose with leucine or
essential amino acids: Effects on
myobrillar protein synthesis at rest and
following resistance exercise in men.
J Physiol 590: 27512765, 2012.
17. Coffey VG, Zhong Z, Shield A, Canny BJ,
Chibalin AV, Zierath JR, and Hawley JA. Early
signaling responses to divergent exercise
stimuli in skeletal muscle from well-trained
humans. FASEB J 20: 190192, 2006.
18. Courthaudon JLJL, Girardet JM,
Campagne S, Rouhier LM, Campagna S,
Effects of Different Dietary Proteins
VOLUME 36 | NUMBER 3 | JUNE 2014
40
Linden G, and Lorient D. Surface active
and emulsifying properties of casein
micelles compared to those of sodium
caseinatebovine micellar casein. Int Dairy
J 9: 411412, 1999.
19. Crozier SJ, Kimball SR, Emmert SW,
Anthony JC, and Jefferson LS. Oral leucine
administration stimulates protein synthesis
in rat skeletal muscle. J Nutr 135: 376
382, 2005.
20. Dangin M, Boirie Y, Garcia-Rodenas C,
Gachon P, Fauquant J, Callier P, Balle` vre O,
and Beaufre` re B. The digestion rate of protein
is an independent regulating factor of
postprandial protein retention. Am J Physiol
Endocrinol Metab 280: E340E348, 2001.
21. Dangin M, Boirie Y, Guillet C, and
Beaufre` re B. Inuence of the protein
digestion rate on protein turnover in young
and elderly subjects. J Nutr 132: 3228S
3233S, 2002.
22. Esmarck B, Andersen J, Olsen S,
Richter EA, Mizuno M, and Kjr M. Timing
of postexercise protein intake is important
for muscle hypertrophy with resistance
training in elderly humans. J Physiol 535:
301311, 2001.
23. FAO. Dietary Protein Quality Evaluation in
Human Nutrition. Report of an FAO Expert
Consultation. Rome, Italy: FAO Food and
Nutrition Paper, 2013.
24. Fouillet H, Mariotti F, Gaudichon C, Bos C,
and Tome D. Peripheral and splanchnic
metabolism of dietary nitrogen are
differently affected by the protein source in
humans as assessed by compartmental
modeling. J Nutr 132: 125133, 2002.
25. Fry AC, Kraemer WJ, Stone MH, Warren B,
Kearney J, Maresh C, Weseman C, and
Fleck S. Endocrine and performance
responses to high volume training and
amino acid supplementation in elite junior
weightlifters. Int J Sport Nutr 3: 306322,
1993.
26. Glynn EL, Fry CS, Drummond MJ,
Timmerman KL, Dhanani S, Volpi E, and
Rasmussen BB. Excess leucine intake
enhances muscle anabolic signaling but
not net protein anabolismin young men and
women. J Nutr 140: 19701976, 2010.
27. Goldberg AL. Mechanisms of growth and
atrophy of skeletal muscle. Muscle Biol
1: 89118, 1972.
28. Goldberg AL, Etlinger J, Goldspink D, and
Jablecki C. Mechanism of work-induced
hypertrophy. Med Sci Sports 7: 185198,
1975.
29. Hartman JW, Tang JE, Wilkinson SB,
Tarnopolsky MA, Lawrence RL,
Fullerton AV, and Phillips SM.
Consumption of fat-free uid milk after
resistance exercise promotes greater lean
mass accretion than does consumption of
soy or carbohydrate in young, novice,
male weightlifters. Am J Clin Nutr 86:
373381, 2007.
30. Herda AA, Herda TJ, Costa PB, Ryan ED,
Stout JR, and Cramer JT. Muscle
performance, size, and safety responses after
eight weeks of resistance training and protein
supplementation: A randomized, double-
blinded, placebo-controlled clinical trial.
J Strength Cond Res 27: 30913100, 2013.
31. Hoffman JR and Falvo MJ. ProteinWhich is
best. J Sports Sci Med 3: 118130, 2004.
32. Joy JM, Lowery RP, Wilson JM, Purpura M,
De Souza EO, Wilson SM, Kalman DS,
Dudeck JE, and Jager R. The effects of 8
weeks of whey or rice protein
supplementation on body composition and
exercise performance. Nutr J 12: 86, 2013.
33. Katsanos CS, Kobayashi H, Shefeld-
Moore M, Aarsland A, and Wolfe RR.
A high proportion of leucine is required for
optimal stimulation of the rate of muscle
protein synthesis by essential amino acids
in the elderly. Am J Physiol Endocrinol
Metab 291: E381E387, 2006.
34. Kelley DE, He J, Menshikova EV, and
Ritov VB. Dysfunction of mitochondria in
human skeletal muscle in type 2 diabetes.
Diabetes 51: 29442950, 2002.
35. Kerksick CM, Rasmussen CJ,
Lancaster SL, Magu B, Smith P, Melton C,
Greenwood M, Almada AL, Earnest CP,
and Kreider RB. The effects of protein and
amino acid supplementation on
performance and training adaptations
during ten weeks of resistance training.
J Strength Cond Res 20: 643653, 2006.
36. Kumar V, Atherton PJ, Selby A, Rankin D,
Williams J, Smith K, Hiscock N, and
Rennie MJ. Muscle protein synthetic
responses to exercise: Effects of age,
volume, and intensity. J Gerontol A Biol
Med Sci 67: 11701177, 2012.
37. Lemon P. Effects of exercise on dietary
protein requirements. Int J Sport Nutr
8: 426447, 1998.
38. Lemon PW and Proctor DN. Protein intake
and athletic performance. Sports Med
12: 313325, 1991.
39. Maughan R. Relationship between muscle
strength and muscle cross-sectional area
implications for training. Sports Med
1: 263269, 1984.
40. Moore DR, Robinson MJ, Fry JL, Tang JE,
Glover EI, Wilkinson SB, Prior T,
Tarnopolsky MA, and Phillips SM. Ingested
protein dose response of muscle and
albumin protein synthesis after resistance
exercise in young men. Am J Clin Nutr 89:
161168, 2009.
41. Ogasawara R, Kobayashi K, Tsutaki A,
Lee K, Abe T, Fujita S, Nakazato K, and
Ishii N. mTOR signaling response to
resistance exercise is altered by chronic
resistance training and detraining in
skeletal muscle. J Appl Physiol (1985)
114: 934940, 2013.
42. Ogasawara R, Yasuda T, Sakamaki M,
Ozaki H, and Abe T. Effects of periodic and
continued resistance training on muscle CSA
and strength in previously untrained men. Clin
Physiol Funct Imaging 31: 399404, 2011.
43. Pedersen BK. Muscle as a secretory organ.
Compr Physiol 3: 13371362, 2013.
44. Pennings B, Groen BB, van Dijk JW, de
Lange A, Kiskini A, Kuklinski M, Senden JM,
and van Loon LJ. Minced beef is more
rapidly digested and absorbed than beef
steak, resulting in greater postprandial
protein retention in older men. Am J Clin
Nutr 98: 121128, 2013.
45. Phillips SM. Dietary protein requirements
and adaptive advantages in athletes. Br J
Nutr 108: S158S167, 2012.
46. Phillips SM, Tipton KD, Aarsland A,
Wolf SE, and Wolfe RR. Mixed muscle
protein synthesis and breakdown after
resistance exercise in humans. Am J
Physiol 273: E99E107, 1997.
47. Phillips SM and Van Loon LJ. Dietary
protein for athletes: From requirements to
optimum adaptation. J Sports Sci 29:
S29S38, 2011.
48. Reidy PT, Walker DK, Dickinson JM,
Gundermann DM, Drummond MJ,
Timmerman KL, Fry CS, Borack MS,
Cope MB, Mukherjea R, Jennings K,
Volpi E, and Rasmussen BB. Protein blend
ingestion following resistance exercise
promotes human muscle protein synthesis.
J Nutr 143: 410416, 2013.
49. Reitelseder S, Agergaard J, Doessing S,
Helmark IC, Lund P, Kristensen NB,
Frystyk J, Flyvbjerg A, Schjerling P, and
van Hall G. Whey and casein labeled with
L-[1-13C] leucine and muscle protein
synthesis: Effect of resistance exercise and
protein ingestion. Am J Physiol Endocrinol
Metab 300: E231E242, 2011.
50. Rodriguez NR, DiMarco NM, and Langley S.
Position of the American dietetic
association, dietitians of Canada, and the
American college of sports medicine:
Nutrition and athletic performance. J Am
Diet Assoc 109: 509527, 2009.
51. Smith K, Reynolds N, Downie S, Patel A,
and Rennie MJ. Effects of ooding amino
Strength and Conditioning Journal | www.nsca-scj.com
41
acids on incorporation of labeled amino
acids into human muscle protein. Am J
Physiol 275: E73E78, 1998.
52. Snijders T, Verdijk LB, and Van Loon L. The
impact of sarcopenia and exercise training
on skeletal muscle satellite cells. Ageing
Res Rev 8: 328338, 2009.
53. Stark M, Lukaszuk J, Prawitz A, and
Salacinski A. Protein timing and its effects
on muscular hypertrophy and strength in
individuals engaged in weight-training. J Int
Soc Sports Nutr 9: 18, 2012.
54. Stepto N, Coffey V, Carey A,
Ponnampalam A, Canny B, Powell D, and
Hawley J. Global gene expression in
skeletal muscle from well-trained strength
and endurance athletes. Med Sci Sports
Exerc 41: 546565, 2009.
55. Symons TB, Shefeld-Moore M, Wolfe RR,
and Paddon-Jones D. A moderate serving
of high-quality protein maximally stimulates
skeletal muscle protein synthesis in young
and elderly subjects. J Am Diet Assoc 109:
15821586, 2009.
56. Tang JE, Moore DR, Kujbida GW,
Tarnopolsky MA, and Phillips SM.
Ingestion of whey hydrolysate, casein, or
soy protein isolate: Effects on mixed
muscle protein synthesis at rest and
following resistance exercise in young
men. J Appl Physiol (1985) 107: 987
992, 2009.
57. Tang JE and Phillips SM. Maximizing
muscle protein anabolism: The role of
protein quality. Curr Opin Clin Nutr Metab
Care 12: 6671, 2009.
58. Tipton KD, Borsheim E, Wolf SE,
Sanford AP, and Wolfe RR. Acute
response of net muscle protein balance
reects 24-h balance after exercise and
amino acid ingestion. Am J Physiol
Endocrinol Metab 284: E76E89, 2003.
59. Tipton KD, Elliott TA, Cree MG, Wolf SE,
Sanford AP, and Wolfe RR. Ingestion of
casein and whey proteins result in muscle
anabolism after resistance exercise. Med
Sci Sports Exerc 36: 20732081, 2004.
60. Tipton KD, Ferrando AA, Phillips SM,
Doyle D, and Wolfe RR. Postexercise net
protein synthesis in human muscle from
orally administered amino acids. Am J
Physiol 276: E628E634, 1999.
61. Verdijk LB, Jonkers RA, Gleeson BG,
Beelen M, Meijer K, Savelberg HH,
Wodzig WK, Dendale P, and van Loon LJ.
Protein supplementation before and after
exercise does not further augment skeletal
muscle hypertrophy after resistance
training in elderly men. Am J Clin Nutr 89:
608616, 2009.
62. Verhoeven S, Vanschoonbeek K,
Verdijk LB, Koopman R, Wodzig WK,
Dendale P, and van Loon LJ. Long-term
leucine supplementation does not increase
muscle mass or strength in healthy elderly
men. Am J Clin Nutr 89: 14681475,
2009.
63. Wilborn CD, Taylor LW, Outlaw J,
Williams L, Campbell B, Foster CA, Smith-
Ryan A, Urbina S, and Hayward S. The
effects of pre- and post-exercise whey vs.
casein protein consumption on body
composition and performance measures in
collegiate female athletes. J Sports Sci
Med 12: 7479, 2013.
64. Wilkinson SB, Tarnopolsky MA,
MacDonald MJ, MacDonald JR,
Armstrong D, and Phillips SM.
Consumption of uid skim milk promotes
greater muscle protein accretion after
resistance exercise than does consumption
of an isonitrogenous and isoenergetic soy-
protein beverage. Am J Clin Nutr 85:
10311040, 2007.
65. Yang Y, Churchward-Venne TA, Burd NA,
Breen L, Tarnopolsky MA, and
Phillips SM. Myobrillar protein synthesis
following ingestion of soy protein isolate
at rest and after resistance exercise in
elderly men. Nutr Metab (Lond) 9: 57,
2012.
Effects of Different Dietary Proteins
VOLUME 36 | NUMBER 3 | JUNE 2014
42