CHAP T=tr

10

Hemoglobin Metabolism

to7

Terminus

{'

glg -

TYR

LYS

'IPHE GLU,.

120

lIS
GLY
t

G
ASf{

vA|T ynsffi Hrsuu

110

&e"-_--,-: "-.

cys 'ffir,,

PHE

-r-a
VAI

*Terminus

n,,

--@

Lys

HIE.
l

$u
THR

619 tHaG n,' TH; Piir r-TS"-' @@

LEU

tr90\ A|-l '

@

ssp
cYS

eru J"- Pno

1....

so

A$N
ASP

:,:)"histidine,
LEU

lunf
ALA.ViL.
ALA

4
'

-lsER

Hts E7 (Distal histidine residue)

i'--'THR

rgr---.-@,
r-Ys

-t- GLY

g'

20

LYS

ALA
66

Close spatial contact

tti'W' PBO.

tY$

D vnrJMrr *sd.r I ASp 40

,::.-i
GLU

q,. lffi
I

56

11.16 PRO

6e

oLY-AsP-r-Eu-ssa

--i. SchrocierWA: Ney,,Aspects of the Structure, Functon, and Synthesrs of

Fxample of a B globin chain. lt is a polypept de chain shouring helical and nonhelical segments. (FrOm Hemoglobins. Boca Raion,
ed,

trress,.]971, modifiecl from Stamatoyannopoulous G: The Molecular Basts of Bl00d Diseases,2nd Saunders, l994.)

.:,ii' of

llb,

also calied

tetrarneric

non-o 1 bonds also contribute to the stabilitv of the structure

(Fig. 10'4)."'i

ir noiecule. The complete IIb rnole' . hen-re groups attached to four pol-vmolecules of oxygtrt"t. lt is chains and lwo non-cr globitr ,.;r has a lretne gloup attached. I]ach -: ..i carrying one rrolecule of oxyrge n. -, c .1, non-o 2 climeric bonds hold

rrrn' lbur

rin

BIOSYNTHESIS

fi**:c
Biosynthesis of heme occurs in the bone marrow in tlie rnitochondria and cfioplasm of'the erythroryle precursors frcrtlr the

'

:n.

'Ietramr'ic

1, non-o- 2 and

g 2,

pronoruoblast/proerythroblast

to the

reticuloryte- Matue

108

PABT

Il

Hematopoiesis

t. ". '.,
. ,1,.t,,, ,
::

".,....]

r,'ll,t,... ...,'.. ::,.'._l*,: ::! : :i_ I 1..... : ...

ftofl-0,1

Complete Hb molecule. Heme is suspended betri,ieen the E anC F hellces 0f the p0lypeptide chatn, Pink represents a, 1 \lef4 artd a. 2 \itghil, yellori represents nan'a. 2 (lell) and non'

fill;r* ill-,{

a.1 lright)

ery1hroc.\"tes cannot rnake Hb because tl-re.v lose their niitochondria ancl the capabilit-v of using tl're tricarbox-vlic acid o'cle

genes .rre

linked on chromosome 11. h'r the human

gc.-

ll('cc\\Ara ior Hb:r ntitesis.

Heme biosynthesis begins n'jth tlre conde nsation trl glvcine and surcinyl coenzvme A (Co,'\) catal-vzed bv aminolevulinate synthase (AL,\S) to form anrinoler.'r,rlinic acicl (AI-A). AI-A dehvdratase (AIA deh.vdrase/porphobi I in ogen s.vnlhase ) in the prsence of AL,\ catalvzes the tbrn-ration of porphobilinogen. Porpl-robilinogen deaminase, also knor,vn as hyclrox,vmethyl-

there is one cop,v of each globin gene per chronaticl for . oi tr.vo genes per person lvith tl-re exception of s. and ^'

oithe o and lgerres perr chrortl:rtid foi a tfour genes per person. lhe prorlr.rction of the globin ch:rins takes place fro:: pronornroblast to the reticttlocvte.* lhc globiir Proteii-.'
arc 1$ro ct-rpies

bilane synthase, in the presence of porphobilinogen catal1'zg5 the formatiort of hyclrox,vl methylbilane. This pathrva-v cor-r' tinr"res until, in the final step of production of heme, Fe-= combines with protoporphvrin lX in the presence of ferrocheiatase/ herrrr synth.rse to tnakt' hemc (l ig. iU l ) '' '[iansferrin, a plasma prolein, carries iron in the l'erric (l'eJ') form to developing tlllCs. lron goes throulh the fl[]C ntenrbrane to the nitocliondria aud is uniled rvitir protctporph.vrirr IX to make heme. Fleme ieaves the mitochondria and is joined to the giobill chains in thc cyioplasm.

made via transcrip{ion of the genetic cnde lct lness: ribonucleic acid (ri-rRNA) and translation of ntRNA t: globin pol-vpeptide chain. A slight excess of u-globin mR'.'

nr

present in erythroblasts; hol ,evet p-globin mRN,\ is tran: ; more elficientl,v rhan cl"-globin nfu\A. This results 1rr s : '1 single chains svnthesized in approximarel-v ec1ua1 alnoi : Whcn slrnthesized, the chains are releasecl from the ribosc :

in tire cvloplasm.t

Six structurai g,enes control the s1'nthesis of the six globin chains. s and ( genes ate on chromosonle 16; T, 0, E, and e

*{*bi*

S*rx*g 1*f:! 3!s** *:l.r[y Afier their release lrom ribosomes, each glcbin chain bin.-' a heme molccule antl tltev pair off. r\u s, cl-rait-t and a tlr chain cornbine tc) form heterodir-ners.'l-wo heterodimers s: taneousl-v combiire to lbrm tetramers. The tetrameric tr,p .a.p, bonds alsr:r contribute io th st.:bilitw of the strrrcrut c. completes the llb n'rolecule (Fig. 10-5)."'7

CHAPTEP

10

Hemoglobin Metabolism

109

- .r:rion of two tx and two B chains along r,r';th the r :::iecules forms Hb A. This is the predominant Hb .-:l.rral life. l'lb A, contains two G and two 6 chains.6 - -.: rre inefficiently expressecl; only small amounts of i{b A. -. :,und in the RBCs. Ifb F contains two cx and two y chains. . jults, Fib F is not distributecl evenlv among the RBCs; it is ,
.(nt in
a few RBCs called

.,.l

rails.:

,ee various globin chains of l-lb differ in the charge per - .ecule. In the procedure of Hb electrophoresis under the - .:ence of an electric field, Hbs exhibit different nrobilities, :rrirg differentiation of subtype. lt mav be necessary to use lrent assay procedures (support rnedia, buffer, pH) for
-efinitive idenrification (Chaprer 26).

The Hb composition in the erythroryte differs, clependinpi on gestation or postnatal age. This change is due to changes in the activation and inactivation or sr,r'itching of the globin genes, progressing from the ( to the c{. gene on chrornosome 1(r and from the e to the 1, 6, or B genes on chromosorne ll. The ( and t genes normallv appear only during the first 3 months of embryonic development. These t.r,r'o chains in addition to the d and l chains are constiruenrs of errbryonic Hbs (Fig. 10-6). Ar birth, Hb Ir is the predominanr Hb. Norrnal adult flb is predonrinately Hb A (urB.) with small amounrs of Hb A, (a.6.) and Hb It (cx.,yr).t'lable 10-2 presents adull reference intenais.

Mitochondrion
Glycine ALA synthase

Cytosol

+>

ALA
I v

Succinyl CoA

ALA d"hudr"t"

Porphobilinogen (PBG)
I

I v
tI

PBG deaminase

Hydroxy methylbilane

UioPorPhyrinogen lll synthase

Uroporphyrinogen lll

Protoporphyringogen
IX

.-Toxrdase

Uroporphyrinogen decarboxylase
ll
I

Coproporphyrinogen

Protoporphvrinoqen
-l oxroase

Coproporphyrinogen
Y
I

Protoporphyrin lX
Fe+2

Ferrochelatase
Heme

polypeptide

chain I

Globin non-u polypeptide chain

@@@@

- r'

Ribosomes I t' I

r;}"i

@-'J_ n t;rFY
F4ry*
Flgure 10-5
Hb assembly

*'M

'lt" non-{{' tetramer

110

PAFT

II

Hematopoiesis
tlr iL
lL

Months

rllull

]ill

llll L"l'

rr

illlll
:ilLnltl

Site of blood cell

production
lil

fr[j-

;::

Figure 1i)-S

Timeline of globin chain produc-

A
50 40
Globin cnarn

{illlN {n'-:lil#
lill{lllll$ll$ il,i*,

tion from intrauterine life to adulthood,

irilllllmtrilllllillr
iiilltlll

"Fl'r

synthesis % of tolal 20
10

Weeks 0 B

10

20

30

lntrauterine lrfe

0 Birth

10

2a

lnfant
lll

IIb A

also has minor amounts

of ilb that have been

been added to the N terminal valine of the

{J

chairr. Normallr'

modi{ied after translation (post-trarrslation). llb A, is forraed b_v postsvnthetic, nonenzymatic reactions of various sugars rvith an-rino groups of tlre globin chains, resulting in glycated Hb. -l'he most common one is Hb A,., in wlrich glucose has

about

49lo

to

6yo

of ilb A is in the A,. form. ln uncontrollec

diabetes n'rellitus, the amount of A,. is increased. C)ther post

rmr
I llru,

translational modifications seen to be of iittle importance.:

REGULATION

l{ervle

TABLg'10**
lntraut*nin*

Normal Hemoglobins

Ear y embryogenesis (product of the

4z+Ez
Cl2

Gower-1

yolk sac erythroblasts) Begins rn early embryogenesrs; peaks

E2 "{z

Gower-2 Portland
F

Regulation of herre production occurs in the heme productior pathway. 'Ihe key rate-limiting step in lreme synthesis is th. initial reaction of gl-vcine and succinyl-CoA to forrn AL\ catal-vzed by.\LAS. Transcription of the Al"\S gene is irrhibitei by heme, rthich leads to a decrease in heme production (: negative feedback rnechanism). C)ther enzymes iir tire herrt

giilrl(lillilrq$
r
lll1

utll

nr
1

1l

(z +
dr,"ng midgeslat 01 and begins

az+\z

pathway inhibited bi' heme are AIA dehydrase and por' phobilinogen deaminase/hy'droxymeth_vlbilane synthase. Ar increased demand for heme woulcl induce an increased svr.rthesis of AIAS.e
Research suggests
pla-vs a regulatorir

llr ill

,l

tl,$
iilt*

;p
uP

r1 v

1'jp6li6o lue!rql vv'v rr hcfnrp u,' dh h r' v

vvu " 'v

Birth
az +
"{z F,

that ferrochelatase (heme synthase) alsc role in I'reme biosvnthesis. A negative feeti-

t,ri lt
lttL.r

60-90%

0z+Fz

A,10-40%

back mechanisnr by heme or substrate inhibition by protoporphylin IX is believed to inhibit the ferrochelatase/heme
synthase enzvme."

1r

tri

Itr'
i

li l
lllill

r
lluf

Adulthond
az +'{z
F, <1-24/a

&!cbin
Clobin production is regulated by the rate at which tl.re DNA i: transcribed to nRNA. The amount of tl're specific globin' s_vnthesized is proportional in ge1-rerirl to the content of thei:

lill

02+62
(I:: + 0z

A?, <3,5%

rilll I '

A, >95%

illlll ti

CHAPTEF
:,vidual globin mILNAs. Heme (in the form of hernin-the - oxidation product of heme) is important in corlrrolling .. rate of globin synthsis in intact rericulocytes and varicrus ..i-fiee syslems, and in its absence, polvribosomes disaggre:..,e. A balanced globin chain and heme synthesis are irrpor, rt because excess components of Hb (unpaired chains, .:toporphvrin, and iron) can decrease RBC survival. The .:d result is that nonlal mature RBCs contain onlv complete :: molecules. Pools of free heme or globin chains are

10

Hemoglobin Metabolism

111

:.

80 c o^^ EOU G l

'.nute.7'e

d40 \o

{ett:*gl*rbin r,r svnthesis is nonnally stimulated by tissue hypoxia.

iipoxia causes the kidn.vs to produce increased amounts of .;'.thropoietin, which stirnulates the prodtLction of FIb and : --Cs. Although each laboratorv nust establish its orvn referranges based on their instrurnentation, methodolog-v, and ..rent ;roprrlation, in general, reference intewals for Hb are as
,\\'s:

20
A. Normal

40

60 Po2 (mm Hg)

'-.

Women

4 1 B g/dl (l 40-1 B0 g/L) 2- 1 5 g/dL (1 20 1 50 g/L) Newllorns I6,5-21 ,5 g/dl (l65 21 5 g/L)
I
1

Men

left- caused by J H+ ions, (1pH), J2,3 BPG, lPco2, itemp (+ Hb variants with I 02 affinity) ,C. Shift to right- caused by f H+ ions, (JpH), 12,3 SPG, t Pco2, ltemp (+ Hb variants with ,f 02 affinity)
B. Shift to

leference inten'als for infants ancl childrer-r vary according :il-ferenl age groups. lndividuals living at high altitucles have :rtlv higher levels of Hb as a compensatory mechanism to lide more oxvgen to th tissues in the oxygen-thin air.'l he - as on the inside front cover of this book show reference -.:ls for all .rgc groupr.

$:igur* 1S-?

Oxygen dissociation curve

FUNCfi6N,,r,:' ',, .r , ,,;r'1", , .: : . .esponsibility of Hb is to bind ox,vlen molecules readil;' re lung (requiring a high aflinit-v for ox,vgen), transport
,

Normall_v, a pO, of approximately 27 mm Hg results in orygen saturation of the Hb molecule. If there is a shift of the cun'e to the left, 507o ox,vgen saturation of Hb occurs at a Po, of less than 27 rnm Hg. If there is a shift of the cur'"'e to rhe
50o/o

-:en

.en, and unload oxygen in the tissues (recluirir"rg a low affir"rity). Each of the four heme iron atorns in a IIb

.:cule can reversiblv bind one oxygen molecule, resulting \fgeDetion of Hl., Approxlmately 1.34 mL of oxygen is
.:rt1 bv each

I g ol Hb.

re affinity of Hb for oxygen depends on the partial pres.' of oxygen (pO,), often defined in terms of the amount of .en needed to saturate 50o/o of Hb, called the Pr,, ualue. |be .::onship is described by the owgen dissociation curve of - rihich plots the ox.vgen content of Hb (% oxvgen satu-

r)

r.'ersus

the pO, (Fig. t0-7). The curve is sign'roidal and

lou' Hb affinity for ox1'gen at lolv oxvgen tension ., high affirritr for o\),gcn.lt lrigh oxygen ten\ions. reration among the subunits of 1{b within the tetramers '.:butes to the shape of the curye. F,ather than unclergo ' -itaneous oxTgenaiion or deoxygenation, the state of eacl-r - .' urlit with regard 1c.l the nunber of other units influences --:r binding. fh.rt is, Hb that is completely deoxygenated ,, :itle affinity for orygen, but with each ox,vgen molecule :: bound, the aviditv increases, and ihe Hb molecule ilv becomes fulll' oyyggrlated.t Shifts of the curve to the ' : nght occur if there are changes in the pH of blood. This -- :n the curve as a result of pH is terrned the Bohr eJfect.
--rtes a

right, 50% orTgen saturalion of Hb occr-rrs at a pO. greater tnan 2, mnl Hp. 'l'he reference inten'al fcrr arteriai ox)en saturation is 969/0 to 1009/0. If the oxygen dissociation cune shifts to the left, a patient rvith arterial and venous pO, concentrations in the reference intervals (80-100 n-rm IIg arteri:rl and 30,50 mm IIg venous) rt'ould have a higher o\ygen sarur;tion percent and a higher affiniv for oxygen rhan a patient r,vith a normal curve. With a shift to the right in the cun'e, a lor,r'er oxygen affinity is seen, and Hb does not pick up as n'ruch oxygen in rhe lr-rrrgs but gives off nrore oxygen t.J the tissue than it norrnally r,vould do, as ir.r the presence of Hb S (Chapter 26). Why and holv 2, 3 -bisphosphoglvcerate ( 2, 3 -B PC ), fonn erly called 2,3-diphospl-rogl-vcerate, aff'ects the cxvgen affinitl' of Hb is a complex process. l'he IIb molecule is an allosteric molecule; that is, its function and structure are influenced bv other molecules. In the presence of large amounts of 2,3-BPC, the I Ib molecule changes from a rela-ted (R) oxygenated rnolecule to a tense (T) deoxygenated molecr:le. lr4any conforma, tional changes occLrr in the Hb molecule for ir to bincl to 2,-l-BPC. The T structure is stabilized by salt bridges, which are broken as the molecule swirches into the 11 str-ucture. Wlren the salt blidges are broken, the Hb molecule is able to bind to ox.vgen (|ig. 10-8). Carbon dioxide, hydrogen ions, and chloride ions all decrease the affinity of Hb fbr oxygen bv strength-

ening the salt bridges that lock the molecule into its

"I'

tt2

PART

II

Hematoooiesis

Figure 10-B

Hb noleclla'cl'anges.

Deoxyhemoglobin tense (T) structure

Oxyhemoglobin relaxed (R) structure

ilillllllllr rrltffid

#ilffiril
qfrtIr1l|flilllilllil

conformation. Some abnormal Hbs with a high oxygen afftnitv and low Prn occur as a result of aniino acid subslitr,rtions that lead to loss of bonds that would have stabilized the tetramer in the'i' conforrnation. Without these binding sites, the I{b molecule holds on more tightly to oxygen-hence a higher
oxygen af{inity.7

rvhereas myoglobinuria could be seen ir.r the aforementioned

disorders.T

{nM.
lllll

ry
I

{mrilN

iiffi!

Clinical conditions that produce a shili to the left include a lowered body temperature owing to external causes; multiple transfusions of stored blood with depleted 2,3-BPC; alkalosis; and the presence of methemoglobin, carboxyhemoglobin, or some other Hb variants. Conditions producing a shift of

the cuwe to the right include increased body temperatur,

increased 2,3-BPC concentrations, increased Ho concentration (decreased pl{), and abnormal Hbs with low affinit1, for ory-

gen. Clinical conditions that prodr-rce a right shift include a high fever; acidosis; and conditions that produce hypoxia, such as high akitude, pulmonary insufficienry, congestive heart failure, severe anemia, and cardiac right-to-left shunt.t Oxygen's sigmoidal curve is in contrast to m-voglobin's hyperbolic cuwe (see Fig. 10-7). lvl.vogiobin, which is present in cardiac and skeletal muscle, is an oxygen-binding heme protein. lt has a molecular r'veight of 17,000 D. It exists in a

Hb F (fetal Hb, the primary Hb in cor:d blood) has increased orygen affinity relative to Hb A. Hb Ir has a P-,, of 19 to 21 mm Hg, resuhing in a left shift of the oxv[ien dissociation curve. The fetus has physiologic ability to extract oxygen from the maternal blood supply via llb F. IIb F does not deliver oxygen .to tissues as efficiently as adult IIb, however. To achieve adequate tissue ox.vgenation, the fetus lrlust have more IIb, and the reference value of Hb for newborns is higher than in the postnatal period when adult Hb predominates. A second crucial function of Iib is in transporting car-bon dioxide in the blood. tn the blood, carbon dioxide undergoes a pair of reactions in which carbon dioxide combirres with water to form carbonic acid, and then carbonic acid disassociates

rffi
rffi,rmlilrrd
i{lllllfrr

nm!

iffiIil

ry
il!,

rilfim

ilillt[

r|ltillililillfir

tftlllililllllllfl

lti

ry
r!(fililJll

$ruutrrtltlill-n16

Illtfriiftilllllitiltl

l|lll|itll'l
I

to release Il* and bicarbonate:

CC), + 1{rO

nil

-+ HrCC)j
HCO.-

:rrLn llllltr

nr

l||tilfilililt!ilililtililililr

HrCf).

+ H'+

rllllllltur

]t tn

$r[ullllliliililr!r,1il]l]rir

The first of these reactions is RBC-facilitated by the enzym

rffillllllllilrsllr
'

monomer form and binds ox.vgen rvith greater affinity tl-ran does Hb. Its hyperbolic curve indicatcs that it releases oxygen only at very low paftial pressures, which means it ls not as
effective as l"lb in releasing oxygen to the tissues at phl,siologic tensior-rs. It is released into the bloodstream u'hen there is

damage to tl're muscle in conditions such as a nyocardial inf'arction, extensive traufira, or severe muscle damage, called rhabdomyolysis. Mi'oglobin normally is flushed out through the kidner', but in the presence of renal failur-e, it ma)i be elevated. Testing for serum nyoglobin may aid in detecting m-vocardial infarction but would be of no value in prtients suspected to have myocardial infarclion who also have extensive trrlulna, rhabdonrvolysis, or renal failure. Elevated m1'oglobin levels give a positive result on the urine Hb dipstick

calbonic anhydrase. fhe FI- fiom the second reaction joins deoxygenated lib. The bicarbonate diffuses across the RBC membrane, and a portion is exchanged witl-r plasma C1-; this i. called the chloride "shift. The plasma bicarbonate travels to the lungs where it is expired. About 70% to 85% of tissue carbon dioxide is processed in this nanner.t0 Approximately 10% tc 2oo/o o{ carbon dioxide binds to the N-terminal amino group of each globin chain as carbaminohemoglobin. Carbaminohernoglobin has a lower affiniry for ox1'gen than does Hb ir
(hc ahserrce of carbolr dioxide.
I

rl|ll lllllill,"
{rlilll

lllillll

lllliitilrrtl

rillllti
lllllftin

iiiilll

"]ll

uli

illtf$lll]llllltllilnil,,

and must be differentiated frorn Hb

in the

urine. llemo-

globinuria wor-rld be seen in intravascular hemolr'tic disorders,

IIb variants whost structure has been r-nodified b-v drugs or envitonmental chern cais. lvlethemoglobin, sulflremoglobin, and carboryhemogir bin are chernicallv modified Hbs.
Chemicallv n-rodified Fibs are acquired

C|IAPT=fl 10
r

HemdglgbinMetabolism

113

l,l':

:r:l

i.: i

1-1

ii

: r..:!;.ir

j t'li

1..,.:it

l'l'l ;', t,l,.r';i

r,r'l

:'i,l

;.",

i;ti

ll r''r,

iiIIi

tletircmoglobin is a form of I{b thar contains iron in the ferric :tate {Fe3'). ,\4etherrroglobin is continuously bsing fonnecl ','ithin erythroq'tes in small anoLrnts because of spontaneous rridatiotr, but it is prer,-enled from accumulating n'ithin the .n'liu'ocytes by the reduction ol'oxidized heme bt' numerous anzvrne svsterns that restlict lhe normal [oncentfation ot

:nethemoglobin ro less rhan toi of total Hb {Chapter 9). \,lethentoglcbin has a brorvnish to bluish color, and it does rLrl rl]vcn io rccl on exposufe to alx]en. Oxidized iron cannol

fiorn the binding of carbon monox, ide to herne ii'on. llb has about 200 tiines nore affinitv for carbon molnxide, and aithr:ugh carbon monoxide combines ra.ith iJb more slorvly than oxygen,'the union is much firrner irnd {he release o1 carbon monoxide is 10,01}0 times'slolver th:rn the relcase of oxr.gen fi'orn deox,virearoglobin. Carbon monoxicle has l:een terrnecl tl-re "silent killer" because n is an odorless arrd colorless gas, and victir-ns mav qr-iicklv become
Carboqrhsnlsglolrfu results

lr1p,'ri,.
Sorne iarbo-xvhemogiobin is produced endogenousiv 'l'he to 0"8%. Exogenous carbon monoxkle is derived fron the exhaust of automobiles anrl fi-om industrial pcrllut;rnts, srich as coal, gas, charcoal burning, and tcbac.o smoke. ln sruokers, levels mav vary/.from 4o/o t<> 2}arc" lixircsure 1o carbon mone--xide may be coincidental, accidental, cl interntional (suicidal). ,\,lany deaths frorn house fires are the result o{'inhaling smoke, flnes, or carbon rnr:noxide.l'r llven ivhen heating systems in the home are properl,v maintained,
reference interval is O.2lo

re oxvgen afEnity ofthe rcnaining heme groups ilcreases.lr il , increased in blood, methmoglobin produces .r shift to the :t1 ir-r tlle oxygen c{issociatiorl .un, and ox,vgen is not delir,ered . ncie ntly to the rissues. if r:-rethemoglobin is greatei rhan 30% : total IIb, patieltls present rvith hypoxia and ryanosis.5,1a Eievated levels of firethernoglolrin zire seen rvher lhere is .rcess production of nethernogl{rbin as a resnlt of presenc - oxiclants {e.g., nitrites) or if there is decreasecl activity oi . r.tllemoglobin redlrctase {usually a genetic dehcienr:v). It also . seen in pat;enls."vhc inherit Hb M disease, which is i:arrsed -. iur itllnorrnalit_v in the siruiture nl Llrc gloLrin fortion of the :-. nolecule (Chapter 26).il \lethenoglobin levels can Lre detectecl by spectrurn .'rbso4";instruments. X.lethemoglobin peaks in the range of 620 to '+0 nm al pl i 7.1. 'lleiltinent of ilcquired melhemogiobinellia

-'ind oxvgen, Lrut u'hen onc or nlorc iron alorrrs have beerr 'rirlized, {he conlbrnrarion of rhe IIb molecule changes, anr,i

accidental poisoning r,r'ith carbon monoxide does occuL.rT 'lcxic effects. such .rs hearlaches and dizziness, mav be present al levcls of i0% ta 75o,/a. I-evels of |l.|of than -5070 may cause
coma and convulsir:ns. Carboxvhemoglol':rin mav be cletected b.v spectral absorption instruments al 541 nm. lt gives blood a

,l

cherrv-recl color, rvhich

is also

in'rparted

poisoning victims.

. :trDoval of the oi'fending

r:

severe carbon rnclnoxicle poisoning, hrrperbar'lc ox1-ge1l lreatments have heen usetl.

In

to the skin of

sutrstance

or adrrjnistraficn of HEMOGLOBIN MEASUREMENT

The reference rnethod used to measllre

,..ri'bic acid or nethviene irlue.is

,-: i,: i r,'':,1.:,i,i :,i:,::

,:.,*emoglobin is a cberntcaliv modifietl Hb fon-necl b.v the .-.ersible oxidation of Hb b.v rertain drugs elnd chemicals,

Iib is the cyanmethe-

'l

moglobiir metleod.rs A lvsing agent plesent in the cl,irnmcthernoglobin reagent is used to fi-ee Hb fr"om the IIB(ls.
Hb conrbines rvith potussirLm ferricv:rnide, r,r'hicl-r is prcsent in the t-r,armelhemogiobin reagent, converting the llb iron fronr the Jerious stat to the {erric state to fornr methemoglobin. f"tetiremoglobin cc.mbines ivith polassiurn cvanide to form the sral:1e pigment, q'anmethemoglobin. Cyanmetherno-

.rs

sulf-onamides, phenacetin, acetarriIide, and phenazopv,

-,ne. |n vitro it is formed by the adclition of"ltydrosen suifitle rlb and has a greenish pignrent. lt is inellective {or ox\.gcn

j1l, anrl patients r.r.ill-r elerrate,:l ievels preser.it r.r.itb q..rnosis. - tlrcmoglobin .;tnltol bc converlecl to ur"rmodified Hb; it
.

,:lspc)r1 {100

tirles less .rlfinity firr orvgern thar-i Lrnrnr.ldifierl

sists

ibr the life ot' the cell. -l'realrne n[ fi)nsists ol inrcid,lncc

nding agent.
''11

.le

qinlrrrr is urctl t,, (lll,-tnli|',Hh hr.re.lrling Llri colnr ch,rng,e speclrophotometricallv rrt 540 nm and cornparing it r.vith a
stalLdar:d i{)hapter: 1aJ. This

ofle

manual ,nethod h;rs been adapted

!,.rlfhernoglobin. sirnilar ro lncthernoglilbin, peaks at 620

.r spef,tral absorption insinirrient.'l-he sull1lernoglobL-r ::ti-.ri cule cloes not shili rvhen c_vanide is adderi, ancl thal ::iLlre .an be useci to distiriguish it from methemoglohin.!

Im

Ior Lrse in ;rutornaled instrr.rnrenls. Il.rnv instruments no\{ use sodium laurvl sulfhte (SLS) to convefi Ilb to SIS-merhe-

moglobin. This ireihocl docs not generat toxic

{t.hapter 39).

b-.151{s

The Hb molecule
orotoporphyrin lX

is

composed

0f f0ur

heme

groups

0f unlike chains. Fach heme molecule c0mbines with One

cfiain.
rJb. contained

Fe2'r and two pairs

polypeptrde p0lypeptide

2.3-BPG produced by the glycolytic pathway facilitates the unlOading 0f 0xygen from Hb in the tissues. The oxygenation curve 0f Hb is sigmoid owing t0 cooperativity
among its subunits, The Bohr effect explains the effect 0f pH 0n the oxygen release mechanism o{ Hb.
The three nornral adult Hbs are HbA. HbA2. and Hb F. HbA. comp0sed 0f two op heterodimers. is the predominant Hb o{ adults.

in RBCs. carries 0xygen to the tissue bound t0 the

ferrous iron in heme. Hb biosynthesis is regutated by hormones.oxygen tension in the {,dneys. and enzymes in the hem synthesis pathway.