``PHYSIOLOGICAL CHEMISTRY the metaproteins, however, the changes in the

original protein molecule are more profound. These derived proteins are
characterized by being soluble in very weak acids and alkalis, but insoluble in
neutral fluids. Themetaproteins have generally been termed albuminates, but
inasmuch as the termination ale signifies a salt it has always been somewhat of a
misnomer.
Two of the principal metaproteins are the acid metaprotein or so called acid
albuminate and the alkali metaprotein or so-called alkali albuminate. They differ
from the native simple proteins principally in being insoluble in sodium chloride
solution and in not being coagulated except when suspended in neutral fluids. Both
forms of metaprotein are precipitated upon the approximate neutralization of their
solutions. They are precipitated by saturating their solutions with ammonium
sulphate, and by sodium chloride also, provided they are dissolved in an acid
solution. Acid metaprotein contains a higher percentage of nitrogen and sulphur
than the alkalimetaprotein from the same source, since some of the nitrogen and
sulphur of the original protein is liberated in the formation of the latter. Because of
this fact, it is impossible to transform an alkali metaprotein into an
acid metaprotein, while it is possible to reverse the process and transform the
acidmetaprotein into the alkali modification.
Experiments on Metaproteins.
ACID METAPROTEIN (ACID ALBUMINATE).
Preparation and Study. " Take 25 grams of hashed lean beef washed free from the
major portion of blood and inorganic matter, and place it in a medium-sized beaker
with 100 c.c. of 0.2 per cent. HC1. Place it on a boiling water-bath for one-half hour,
filter, cool, and divide the filtrate into two parts. Neutralize the first part with
dilute KOH solution, filter off the precipitate of acid metaprotein and make the
following tests: (1) Solubility. " Solubility in the ordinary solvents (see page 27).
(2) Millon's Reaction.
(3) Coagulation Test. " Suspend a little of the metaprotein in water (neutral solution)
and heat to boiling for a few moments. Now add 1-2 drops of KOH solution to the
water and see if themetaprotein is still soluble in dilute alkali. What is the result and
why? (4) Test for Loosely Combined Sulphur (see page 108).
Subject the second part of the original solution to the following tests: (1)
Coagulation Test. " Heat some of the solution to boiling in a testtube. Does it
coagulate? (2) Biuret Test.
PROTEINS
(3) Influence of Protein Precipitants. " Try a few proteinprecipitants such as picric
acid and mercuric chloride. How do the results obtained compare with those from
the experiments on egg albumin? (See page 102).
ALKALI METAPROTEIN (ALKALI ALBUMINATE).

Preparation and Study. " Carefully separate the white from the yolk of a hen's egg
and place the former in an evaporating dish. Add concentrated potassium hydroxide
solution, drop by drop, stirring continuously. The mass gradually thickens and finally
assumes the consistency of jelly. This is solid alkali metaproteinor "Lieberkuhn's
jelly." Do not add an excess of potassium hydroxide or the jelly will dissolve. Cut it
into small pieces, place a cloth or wire gauze over the dish, and by means of
running water wash the pieces free from adherent alkali. Now add a small amount of
water, which forms a weak alkaline solution with the alkali within the pieces, and
dissolve the jelly by gentle heat. Cool the solution and divide it into two parts.
Proceed as follows with the first part: Neutralize with dilute hydrochloric acid, noting
the odor of the liberated hydrogen sulphide as the alkali metaprotein precipitates.
Filter off the precipitate and test as for acid metaprotein, page 116, noting
particularly the sulphur test. How does this test compare with that given by the acid
metaprotein? Make tests on the second part of the solution the same as for acid
metaprotein, page 116.
Coagulated Proteins.
These derived proteins are produced from unaltered protein materials by heat, by
long standing under alcohol, or by the continuous movement of their solutions such
as that produced by rapid stirring or shaking. In particular instances, such as the
formation of fibrin from fibrinogen (see page 195), the coagulation may be produced
by enzyme action. Ordinary soluble proteins after having been transformed into the
coagulated modification are no longer soluble in the ordinary solvents. Upon being
heated in the presence of strong acids or alkalis, coagulated proteins are converted
into metaproteins.
Many proteins coagulate at an approximately fixed temperature under definite
conditions (see pp. 106 and 2 54). This characteristic may be applied to separate
different coagulableproteins from the same solution by fractional coagulation. The
coagulation temperature frequently may serve in a measure to identify proteins in a
manner similar to the melting point or boiling-point of many other organic
substances. The separation of proteins by fractional coagulation is thus analogous
to the separation of volatile substances by means of fractional distillation. This
method of separating proteins is not a satisfactory one, however, inasmuch as
proteins in solution have different effects upon one another and also because of the
fact that the nature of the solvent causes a variation in the temperature at which a
given protein coagulates. The nature of the process involved in the coagulation of
proteins by heat is not well understood, but it is probable that in addition to the
altered arrangement of the component atoms in the molecule, there is a mild
hydrolysis which is accompanied by the liberation of minute amounts of hydrogen,
nitrogen, and sulphur. The presence of a neutral salt or a trace of a mineral acid
may facilitate the coagulation of a protein solution (see page i c6) ,whereas any
appreciable amount of acid or alkali will retard or entirely prevent such coagulation.
It has recently been shown that the coagulation of proteins by heat proceeds in two

stages,1 first, a reaction between the protein and the hot water(denaturation) and
second, an agglutination or separation of the altered protein in particulate form. The
concentration of acid, or hydrogen ion, in the solution influences the coagulation of
proteins, such that the original protein is acted upon less readily by hot water alone
than in the presence of acid. The formation of the coagulum is accompanied by the
disappearance of the free acid from the solution, indicating the formation of a
protein salt. A disturbance of the equilibrium between the hydro lyzedand
unhydrolyzed portions of the protein salt, due to the greater rapidity with which the
unhydrolyzed portion is precipitated, results in the gradual removal of both protein
and acid from the solution. This has been offered as an explanation of the
decreasing acidity.
According to Chick and Martin, the addition of neutral salts to the acid solution of
the salt-free protein to be coagulated results in a. decreased rate of coagulation.
This is due in part to the decrease in the concen- tration of the free acid, which
results from the disturbance of the equilibrium between the protein and acid and
also in part to the direct influence which the salts exert upon the protein. The
presence of neutral salts may under certain circumstances facilitate the coagulation
of proteins by heat.
The temperature at which egg white is coagulated causes a difference in the
appearance of the coagulum.2 Coagulated egg white which has been immersed in
water at a low temperature and then gradually heated to the coagulating
temperature is more translucent and has a bluish color, whereas, egg white which
has been immersed in water heated to a temperature above the coagulating
temperature is creamy-white in color. The varying digestibility, as the result of the
different methods of heating has been discussed in the chapter on Enzymes.
----------------------------------------------------------------------------------------------------------------------------------------Metaproteins are formed by the action of fairly concentrated acids and alkalis on
proteins. These are very soluble in very dilute acids and alkalis but insoluble in
neutral solvents, for example, acid metaproteins and alkali metaproteins
These are hydrolytic productsof albumins and globulins formed by the action of
water or dilute acid or alkali. They are insoluble in water, strong mineral acids, and
all solutions of neutral salts, but are soluble in dilute acid and alkalis in the absence
of any large amount of neutral salt.
These are produced by hydrolysis of natural proteins, by alkalis or prolonged
treatment with acids.