roteins are the ,,,_, abundant orpnk
P molecuk, of the lir"'6 •y,tffll. They occur
in every part of the cell and constitute about
SO% of the cellular dry weight. Proteins form
the fundamental basis of structure and function
of life.
Origin of the word 'protein'
The term protein is derived from a Greek
word proteios, meaning holding the first place.
Berzelius (Swedish chemist) suggested the name
proteins to the group of organic compounds that
are utmost important to life. Mulder (Dutch
chemist) in 1838 used the term proteins for the
high molecular weight nitrogen-rich and most
abundant substances present in animals and
plants.
Func:tlens of proteins
Proteins perform a peat variety
and essential functions in the I
functions 1MY be bloadl
Lt .If,
Struclunl functlMI : Celtaln pt8Mlf11 pe,efolffl
brick and mortar roles and lft9 ;Aiklly
responsible for structure and ••lllh flA l:todr·
These include colllflM and eb l&t found rtt bent
matrix, vascular system and ad.. 011i1f11 ..e
a-keratin present In epidermal tillliel.
Dynamic: functlonl : The dynamic fufteftont
of proteins are more diversified in nature. Thele
include proteins acting at - - , s t . . .,.,,
blood
membrane receptors, storage proteim_
•••
do,,.,, ,-.,.,.,. . . •...,-•dN I\
their function in ~ cont,ol
contraction, respiration etc. Protelnt
dynamic function5 are approp,lllely ,.,Ed..
the . . . . . . , _ _ of cell.
 tom t,lndt to • tide d,afn
llellildlll thtt ' rh• ~ .,t,on • hi hIt different to, each ol
ahow ,,,.,.,n, may 1110 on!A " r pr•"nwd by R ~ound In proC.rlM The am,no
h p u I Ml, Mn, ln ., th 20 ,mlr,o , fdt '1 the Jonlzed fo,m In 1he
nltropn, nn u ntlal II ldt mmtly ""''~1 ,.,';,w,, 8 t,cwe)

n
O '"H" ,. ''"·
toty (rm>1tly
t al o (ln<I out I,,
t In In hloln d11 nnd (ood11
•r• polvmor of ■ mlno acid•
hydrol (with oncrn
h ur ) Id l a nmlno
y o( nll tho
on pro
~,n, 111·11 the polymer, of
ID
lno .1 Id oc.c:ur m nMurc
known ,; 1:indrtrd amino
found In the ,truc:turc of
m rllff mnt forrm of llfc-
I roblol rh11; Is hccnuse of
( the g nellc c1Jdc available
of only 20 ammo adds
ar syn1hcslzcd in the ccll8.
rn I controlled by DNA, the
( tho cell. After the synthe111s of
of the Incorporated ammo acids
u odlflcatlon11 to form their derivatives.
AMINO ACIDS
Amino acids are a group of organic
compound• containing two functional ,,ou,,._
Mrtlno and arboxyl The amino group (-NHz)
11 b11lc whlle the carboxyl group (-COOH) ls
acidic In nature
General •tructur• of amino acid•
The amino acfdt are tarmed •• a-amino acids,
If boCh the carboxyl and amino 1roops are
.u.ched to the same carbon atom, a, depleted
below
hlolm4I(1JI ,y,tr.m
of .,,,ano •cld•
0 P tic •• l•om•r• attachl!d tc, four different
15
If 11 <.nrhon awm etrlc and therefore exhibit,
Hroupt, 11 It a,ymm The amino acids ( ~
or,1lc::1f ltomerltmf ur dlttmct groupt (R, H,
O
gf ydnc) po!JCJ~• Id t,y a-carbon, Thus all the
COO , NHJ) e glycine where R • H) have
amino acids (excePI
optical Isomers. .
of L· and D-amino ac,dt are
The structuret the configuration of L· and
written based ~n shown in Flg.4.1, The
D glyccrafdehy e as d of L-<1,-ammo adds,
protcini, arc compose
c1a11slflcatlon of amino acids
•fferent ways of classifying the
There are dl d he . 1
amino acids based on the structure an 1· c.& mica
'tlonal requirement metabo IC ,ate etc.
nature, nut r, '
A Amino add danification bated on the
,tru~ure : A comprehensive classificationof
amino acids is based on their structure and
chemical nature. Each amino acid is assigned a
3 letter or 1 letter symbol. These symbols are
commonly used to represent the amino acids in
protein structure. The 20 amino acids found in
proteins are divided into seven distinct groups,
In Tabl~ 4.1, the different groups of amino
acids, their symbols and structures are given. The
tallent features of different groups are described
next
 S 45
Ch• pter 4 PROTEINS ANO AMINO ACIO

lno iitlds found In prote ins

fAai
1
4 l ~lru , tur•I cl.iu lflt•t lon of L "••m

Stl'l lduN

G H ?H- COO -
1 --
NH;
2. Alanine All A CH 3 CH- coo -
I +
NH3

H3C
' CH - CH- COO- BrwlChld chlln
3. Valine Val V ,, I
H3C NH;

H3C,
CH - CH 2 - CH-C
I OO"' Brlnchld dllln
4. Leuclne Leu L ,,
H3C NH;

CH 3
' CH ,2
;cH - CH-COO
-
5. lsoleuclne lie
H3C ~;
-----
······--------
················
... · · · · · · · · · · · ················
. ........ .. . ..... - -
. . . . . . . . . . . . . .- u ~ ~

N. Amfno ICldt contllmng hydroxyl (-OH) group

e

Ser s CH 2 CH-COO-
6. Serine i. I +
OH NH3

Thr T H3C - CH- yH -co o-

7. Threonine
6H ~;
y SN llldlr ll'Offllllc
 -
-COO-
8CYllllne Cys C CH ?"4+
SH Nt-f3

CH 2 CH-C00-
1 I +
s NH3 Disulfide
s
tH -CH-coo-
I 2
NH;
Thioelher
CH
2
CH2 CH-COO-
9. Methionine Met M i 1 +
_________ ·- ____ .. --······· S...cH3 ....... ~ -~··· ············-·······-··· ···· ······· .... -· ... ····-········-
IV. Addle amino aclda and their amldel
13 a
ooc CH2 CH-COO- tJ-Carboxyl
i +
10. Aspartic acid Asp D NH3

Amide
11 Alplragine Asn N H2N-~-CH2 9H_;-COC1"
0 NH3

Y 13 a _
y-Carboxyl
12. Glutamic acid Glu E -ooc-c~ cH2 -9H-COO
NH:
Amide
13. Glutamine Gin a
- .... •• •••-•• •• •••• ••• ••-•• •••• •• • •• •• o n••n ••••n ••• .......................... ... ................. u .. uooooouu••••-O'•oa••••••••••u••-•••••••••••••••••••••••••••••••••• •- •

v. ........ ...

14.Lylinl K !·Amino

11. , ......
lmldlZole
 Chapter 4 : PROTEINS ANO AMINO ACIDS
47

Name Symbol
Structure Special aroup p,nent
3 letters r letter
VI. Aromatic amino acJdt

17. Phenylalanine Phe F

f
Benzene or phenyl

18. Tyrosine Tyr y

Phenol

19. Tryptophan Trp w lndole

H
.... ·······-----------~·······················-••·-••························-·-••·••-··-··-------- -
VII. lmlno acid

20. Proline Pro p Pyrrolidine

(Note : R group is shown in red)

1. Amino adds with aliphatic side chains :
These are monoamino monocarboxylic
acids. This group consists of the most
simple amino acids-glycine, alanine,
valine, leucine and isoleucine. The last
three amino acids (Leu, lie, Val) contain
branched aliphatic side chains, hence
they are referred to as branched chain
amino acids.
2. Hydroxyl group containing amino acids :
Serine, threonine and tyrosine are
hydroxyl group containing amino acids.
Tyrosine-being aromatic in nature-is
usually considered under aromatic amino
acids.
3. Sulfur containing amino acids : Cysteine
with sulfhydryl group and methionine
with thioether group are the two amino
acids incorporated during the course of
protein synthesis. Cystine, another
important sulfur containing amino acid, is
formed by condensation of two molecules
of cysteine.
4. Acidic amino acids and their amides :
Aspartic acid and glutamic acids are
dicarboxylic monoamino acids while
asparagine and glutamine are their
respective amide derivatives. All these
four amino acids possess distinct codons
· for their incorporation into proteins.
5. Basic amino acids: The three amino acids
lysine, arginine (with guanidino group)
and histidine (with imidazole ring) are
dibasic monocarboxylic acids. They are
highly basic in character.
6. Aromatic amino acich : Phenylalanine,
tyrosine and tryptophan (with indole ring)
 BIO CH EM IErr J::r v
48
'al or indispensable amino •chis .•
Essen• 'h
The 'amino acid s bodwh,c dann
ot h.
are aro m t, mm o a ,ds B Id th Y an , ther efor e,
und r the1ized by the
hf tidi n m riy al o be on ldcr cd ugh
,ynd to be sup plie d thro 'd the diet are
th, 1 ory nee s. The y are
essential amm o ac, h
7 lmino acids : Pr Im ont 11nfn pyrrol1d1nc ca II ed
requ ired for pro~edr . 'dgrowtl h and
rm I unlq u mfn o acid It has an ce of the ,n 1v1 ua . T .e ten
,unt en an
1mino rou p NH ) ,n r ad of .in amino 111
. acid s fisted belo w are esse ntia l
for
amm o
rou p NH ) found 1n othe r amino acids. hum ans (and also rats) :
h re I rol,n 1s an <X-1mmo acid .. e Vt1lme ' His tidi ne, lsol euc ine,
Argmin , • . .
-
B. Cla ssif icat ion of ami no add s
based on • e Lysine' Met hion ine, Phe nyla
d into 4 Leucm ,
pol arit y . mm o c,ds are c/ass1f1e lanine, Thr eon ine, Try ptop han .
d on thei r pola rity The pola rity '" t l~tter
... " ...,.ni_, b [The cod e A. V. Hl~L, MP., T. T. (firs
ts the func tion al role of amm o acid s in be mem oriz ed
ino acid ) may .
tru ture o f eac h am .
Oth er
to recall esse ntia l amm o acid s.
ami no LM P; PH.
1 Non~polar amino acids : These useful cod es are H. VITTAL ,
ids are also refe rred to as hyd roph obic VlLLMA, TT, PVT TIM HALL
and
on
"at er hatrng). The y hav e no cha rge MATTVILPhLy.J
acid s incl ude d
the R gro up. The amm o The two ami no acid s nam ely argi nine
and
leuc ine,
m this gro up are - alan ine, hist idin e can be syn thes ized by
adu lts
nyl-
,sol euc me, valm e, met hion ine, phe and not by grow ing chil dren , hen ce
thes e
alan ine tryp toph an and prol ine. ami no
are con side red as sem i-es sen tial
on 'R' acids (rem emb er Ah, to reca ll). Thu s,
8
2. Pol ar ami no acid s with no cha rge
, carr y l whi le
gro up : The se ami no acid s, as such ami no acid s are abs olut ely esse ntia
eve r
no cha rge on the 'R' gro up. The y how 2 are sem i-es sent ial.
roxyl,
pos sess gro ups suc h as hyd am ino
icip ate in 2. Non-essential or disp ens able
sulf hyd ryl and ami de and part acids : The bod y can syn thes ize abo ut
10
cture.
hyd rog en bon ding of prot ein stru needs,
glyc ine (wh ere ami no acid s to mee t the biol ogic al
The sim ple ami no acid in the
gory . hen ce they nee d not be con sum ed
R = H) is also con side red in this cate ine, seri ne,
are - diet. The se are- -gly cine , alan
The am ino acid s in this grou p ate,
cyst eine, cyst eine , aspa rtat e, asp arag ine, glutam
gly cine , seri ne, thre onin e, .
. glut ami ne, tyro sine and pro line
glu tam ine, asp arag ine and tyrosine
group: D. Amino acid classification based
on their
3. Polar amino acids with positive 'R' of am ino
The thre e ami no acid s lysine, argi
nine metabolic fate : The carb on ske leto n
the syn thes is
and hist idin e are incl ude d in this
grou p. acids can serv e as a prec urso r for
nic) or bot h.
group : of gluc ose (gly cog enic ) or fat (ket oge
4. Polar amino acids with negative 'R' no acid s are
aci ds- From met abo lic view poin t, ami
The dica rbo xy/ ic mon oam ino Re kr
aspart.ic acid and glut ami c acid are divided into thre e gro ups (for deta ils,
Chapter 15).
con side red in this gro up.
se am ino
acids : 1. Glycogenic amino acids : The
C. Nutritional cla,sification of amino for the
are requ ired acid s can serv e as pre cur sors
The twe nty ami no acids (Table 4.1) e.g.
eins , besi des for~ atio n of gluc ose or gly cog en.
for the syn thes is of vari ety of prot e etc.
r, all these 20 alan ine, aspa rtat e, glyc ine, met hio nin
oth er bio log ica l fun ctio ns. How eve
the diet . Based 2 · ketogenic amino acid s : Fat can be
am ino acid s nee d not be taken in
no acid s are s. Two
on the nut ritio nal requirements, ami syn_th esiz ed from the se am ino acid
ial and non - ami no ac1'ds Ieuc ,ne · •
grouped into two clas ses -ess ent and lysi ne are
essential. exc lusi vely ketogenic.
 Chapt
- er 4 : PFIOTEINS
ANO AMINO ACIOS
49
3. Clycogenlc and keto .
The four amino " . genie amino acid1 t
alanine, tryprop~1:1~s lsolPuc1nc, phunyl 3 Taste I Armrw acids may b sweet (Gly,
cursors for synrh • , tyro inc arc pre Al 1, Vnl), lastc I !IS (I cu) or bitter (Arg, lie).
fat. cs1s <>I gluc(Jse os wdl a M,m,,sodlum glutamltlc (MSG, ajinomuto1 1
u ul 1111 n llnvmlng g, nt in food lndu try, and
Sclonoc t . <.:l11nc c lood to lnc.rca c Ul le and O vor In
Ys eano-th 1
'- m1e indlv1duals Intolerant to MSc;, Chinese
e 21st amino acid
As already srat"'d 2 restaurant syndrome (hrief and reversible flu
commonly found ;n c ; _o amino at:1cJs ,ire llke syrnploms) is observed.
21st amino acid , J>I ote1ns. In ri cent yer1rs .1 •I. Optical properties : All the ammo acids
1nme y s ,1 •
added. It 1s r10 d 1; enocys11•1n1• has been
' un
enzymes/proteins (
at the r
ac: ive SIies of certain
cxc,•111 glycine possess 011I1cal isomers due to the
prcscnc c of asymmelric carbon atom. Some
. se1enoprot · )
th tone peroxidase I . ems. e.g. gluta- ,1mino ,1cids illso have a second c1symmetric
dinase thiored . ' 8 ycinc rcduc.tase, 5'-deio- carbon e.g. isoleucinc, threonine. The structure
'
an unusual
0
xin
. reductas" s
. • e. eIcnocyste1ne . .1s
of L and D-amino acids m compc1nson with
< amino acid c t · •
element selenium in I on a1ning the trace glyceraldehycle has been given (See Fig.4. 1,.
cysteine. P ace of the sulfur atom of
5. Amino acids as ampholytes: Amino acids
contain both acidic (-COOHJ and basic
CH2-CH-coo-
l CH2-CH-coo- (-NH 2) groups. They can donate a proton or
SH NH l I
3 SeH NH; accept a proton, hence amino acids are regarded
Cy•telne as ampholytes.
Selenocystelne
ln:orpora_tion of selenocysteine into the Zwitterion or dipolar ion : The name zwitter
proteins during translation is carried out by the is derived from the German word which means
codon_ namely UGA. It is interesting to note that hybrid. Zwitter ion (or dipolar ion) is a hybrid
UGA rs normally a stop codon that terminates molecule containing positive and negative ionic
protein biosynthesis. Another unique feature is groups.
that selenocysteine is enzymatically generated The amino acids rarely exist in a neutral form
from serine directly on the tRNA (selenocysteine- with free carboxylic (-COOH) and free amino
tRNA), and then incorporated into proteins. (-NH 2) groups. In strongly acidic pH (low pH),
the amino acid is positively charged (cation)
Pyrrolysine - the 22nd amino acidl : In the
while in strongly alkaline pH (high pH), it is
year 2002, some researchers have described yet
negatively charged (anion). Each amino acid has
another amino acid namely pyrrolysine as the a characteristic pH (e.g. leucine, pH 6.0) at
22nd amino acid present in protein. The stop which it carries both positive and negative
codon UAG can code for pyrrolysine. charges and exists as zwitterion (fig.4.2).
lsoelectric pH (symbol pl) is defined as the
Properties of amino acids pH at which a molecule exists as a zwitterion or
The amino acids differ in their physico- dipolar ion and carries no net charge. Thus, the
chemical properties which ultimately determine molecule is electrically neutral.
the characteristics of proteins. The pl value can be calculated by taking the
average pKa values corresponding to the ionizable
A. Physical properties groups. For instance, leucine has two ionizable
groups, and its pl can be calculated as follows.
1. Solubility: Most of the amino acids are
usually soluble in water and insoluble in organic pK,(cooH)+PK2(NH;)
solvents. pH=-~-------
2
2. Melting points: Amino acids generally pl= 2.4 +9.6 = 6.0
melt at higher temperatures, often above 200°C. 2
 Cha pter 4 : PROTEINS
~C2!1D~
A
S!_~
l 52 ~o
~:~~v~~- - - - - - - - - -_!
!51 ,;.
-...;..-...:..:..:_::~~~A~N~O~A;t_M~IN~O~A

Reactions due to -NH2 8roup odd ilrt• Vt ry lmporl 111 for J1'0I I
l1111Lllu11 5t•/,.c.'1cd / gt~
4. The nm/no grou I>s I>cl i.wc ,1s li,1, f' i111d
combine . I I11 1 1l'lllld iJr ,
salts ( NI ~clII ac,ds (C' H- IICI) lu (11,n,
protein in
~ ). • Cnll,,~Cll rho ,no
rm yprallne , ff.tfq
lllilll lltl,I IS- lOlll cl Ill 1
5. Reaction w'th
"d
I •h .
nm ydrm : The u-01111 no
ac, li rc.1tt \\llh ni11h}dr111 to fu, 111 , 1
S•hy1/nuyly1/ne. ~tt
purple' blue or PIil k colour lumplt1 x fc,unc/ rn as ocratron
gd,
(Ruhcmann's purplt•). with DNA con10111 m.iny mcrhylal.
acids.
µhosphoryh11cd ur .iccttltitcd amino
Amino nr.id + N111h> drin---..+ Keio add
t
NH3 + CO2 + Hydri11d,111tin • y-C.1tboxyglut.1mlc acid IS founcr 1n ?tefn
1irdrindantin + NH3 + Ninhrdrin ~ plasma prolems involved in bloocf clotting.
RuhcmJnn'~ purple two
• Cysline is formed by combination of
as
cysteines. Cystine is also considered
Nrnh> drin reaction b effectively used for
the quantitative determination of amin
o derived amino acid.
acid:; and proteins . (Note : Praline and B. Non-protein amino acids : These ami
no
hvdroxyproline give yellow colour with acids, although never found in proteins, perf
orm
ninhydrin). func tions. The y
several biologically important
s. A
no may be either a-or non-a-amino acid
6. Colour reactions of amino acids: Ami their
selected list of these amino acids along with
acids can be identified by specific colour
reactions (See Table 4.3) . functions is given in Table 4.2.
C. D-Amino acids : The vast maj
ority of
no
7. Transamination : Transfer of an ami and plan ts are
group from an amino acid to a keto acid amino acids isolated from animals
also
to form a new amino acid is a very of L-category. Certain D-amino acids are
-D,
found in the antibiotics (actinomycin
important reaction in amino acid e and
metabolism (details given in Chapter 15). valinomycin, gramicidin-5). O-serin
e. D-
D-aspartate are found in brain tissu
s pres ent in
8. Oxidative deamination : The amino acid Glutamic acid and D-alanine are
undergo oxidative deamination to liberate bacterial cell walls. S:f '}.,
free ammonia (Refer Chapter 15). ~1--5; 3 : 31t
Amino aci ds use ful as dru gs
NON-STANDARD AMINO ACIDS that
There a certain non-standard amino acids
s
Besides the 20 standard amino acid are used as drugs.
protein
(described above) present in the (D-dimethylglycine), a
structure, there are several other amino
acids • D-Penicillamine
the
which are biologically important. These
include metabolite of penicillin, is employed in
This is
the amino acid derivatives found in prot
eins, chelation therapy of Wilson's disease.
ely
non-pro!ein amino acids performing spec
ialized possible since O-penicillamine can effectiv
functions and the D-amino acids. chelate copper.
sis, and
A. Amino acid derivatives in protein
s : The • N-Acetylcysteine is used in cystic fibro
tion
20 standard amino acids can be incorpor
ated chronic renal insufficiency, as it can func
ersa l
into proteins due to the presence of univ as an antioxidant.
acids
genetic code. Some of these amrno ed to
protein • Gabapentin (y-aminobutyrate link
undergo specific modification after the ant.
of amino cyclohexane) is used as an anticonvuls
synthesis occurs. These derivatives
 BIOCHEMISTRY
52 o~tSill
tmporl•nl no•. p,01,tn •mlno ,cld• ••••• wllh the I• fun,Uons
Tua.1 4.2 A ~~klt~d lbt of
function(•>

o:-Am no adds
thesis of urea.
lntenned1ates In the blosyn

ArgU10Succ1mc aetd
• ed from tyrosine.
Thyroid honnones denv

;-Aoeno:syur1emIon n Methyl donor in biological system. . A risk factor for coronary heart
. . metabOhsm.
lntennediate in methionine .
diseases . rt te and meth1om • ·ne metabolisms.
Intermediate in threonine, aspa a for melanin pigment.
as a precursor
1nyaroxy phenylalan ne (DOPA) A neurotransmitter, serves . .e
d excreted in unn
e Derived from muscle an . f rti'lized eggs, and acts as an
• acid found 1n e
Sulfur containing amino
antioxidant
Azase ne
I • Non-a-am no acids
~ e
Component of vitamin pantothenic_acid and coenzyme A
Aminotsobutyric acid End product of pyrimidine metabolism. . .
y-Am obutync aad (GABA) A neurotransmitter produced from glutam1c acid
6-Amino evu n c acid (ALA) Intermediate in the synthesis of porphyrin (finally heme)
Taurme Found in association with bile acids.

polypeptide chains referred to as subunits. The

STRUCTURE OF PROTEINS
Proteins are the polymers of L-a.-amino acids.
The structure of proteins is rather complex which
can be divided into 4 levels of organization
Fig.4.4) :
1. Primary structure : The linear sequence of
ammo acids forming the backbone of proteins
poJypept,des).
2. Secondary structure: The spatial
arrangement of protein by twisting of the
polypeptide chain.
3 Tertiary structure : The three dimensional
structure of a functional protein.
4 . Quaternary structure : Some of the
proteins are composed of two or more
spatial arrangement of these subunits is known
as quaternary structure.
[The structural hierarchy of proteins is
comparable with the structure o( a building. The
amino acids may be considered as the bricks,
the wall as the primary structure, the twists in a
wall as the secondary structure, a full-fledged
self-contained room as the tertiary structure. A
building with similar and dissimilar rooms will
be the quaternary structure).
The t~rm protein is generally used for a
po~ypept1de containing more than SO amino
acids. In recent ye 5 h
have been using . ar , owever, some authors
'pol •.1-1
number of . . !~Pt•uc even if the
prefer to a;smo acid~ is a few hundreds. They
e protein to b
polypeptide chains With an assem ly of
quaternary structure.
 53
Chapter 4 : PROTEINS ANO AMINO ACIDS

'
Primary
1tructure
Secondary
ltructure
Tertiary
1tructure
,.,,,...,,,.,1on...,,.ry
Quaternary
1tructure
Fig. 4.4 : Diagrammatic
(,._: The four aubunlta of two l)J»S In
of
....,.).
protB/n sttucture

PRIMARY STRUCTURE OF PROTEIN double bond in character. It generally exists in

trans configuration. Both -C=O and -NH
Each protein has a unique sequence of amino groups of peptide bonds are polar and are
acids which is determined by the genes involved in hydrogen bond formation.
contained in DNA. The primary structure of a
protein is largely responsible for its function. A Writing of peptide structures: Conventionally,
vast majority of genetic diseases are due to the peptide chains are written with the free amino
abnormalities in the amino acid sequences of end (N-terminal residue) at the left, and the free
proteins i.e. changes associated with primary carboxyl end (C-terminal residue) at the right. The
structure of protein. amino acid sequence is read from N-terminal end
The amino acid composition of a protein to C-terminal end. Incidentally, the protein
determines its physical and chemical properties. biosynthesis also starts from the N-terminal amino
acid. ·
Peptide bond
The amino acids are held together in a protein
by covalent peptide bonds or linkages. These
H H
bonds are rather strong and serve as the + I + I
cementing material between the individual H:3N-c-coo- +
I
H/4-9-coo-
amino acids (considered as bricks). R1 ~
formation of a peptide bond : When the Amlnoacld1 Amlnoacld2
amino group of an amino acid combines with ~o
the carboxyl group of another amino acid, a
peptide bond is formed (Fig.4.5). Note that a H H
dipeptide will have two amino acids and one + I I
peptide (not two) bond. Peptides containing ~N-C-CO--NH-c -coo-
1 I
more than 10 amino acids (decapeptide) are R1 ~
referred to as polypeptides. Dlplplkll

Characteristics of peptide bond, : The .. - ~.,

' ., • ~ ,,;. '.. ~ ~' I • 4" •

peptide bond is rigid and planar with partial

 54

Shorth.ind
to rNd peptides : The H3N-glutamatt-CYatelne-glycln&-COO
arnino acids tn a peptide or protein
E C - G
are l'Pnresented by the 3-letter or one
·-,.. , · the
letter abbrev1at1on This is . Glu Cys - Gly
~ I 'Sltorthand to write proteins.
GlutamyI - cystelnyl - glycine
Naming of peptides : For na~ing
peptides the amino acid suffixes
__ ~-.-u..-- 0-:-,,-ym=bol::,-:l;:n:f'flf)::,.~.,,~tlng~;.:~::::::---
Rg. 4.I ·tide with 3 amino aclda and M o ~
- - (glycme) -an (tryptophan), -~te (Nol9: A trlpepW Is on the ,.,, whl,. ff'H -COO- le Cln ,_
(glutamate) are changed to -yl ~' th shown; F,w-N 3
the exception of C-terminal ammo N-
. •
acid. Thus a tnpept,de compo sed of an •nal 2 _ Degradation of protein or P<>lyPePticfe
terminal glutamate, a cysteine and a C-~erm, into smaller fragments.
glycine is called glutamyl-cysteinyl-glycme. 3 _ Determination of the amino acid sequell<:e.
In the Fig.4.6, the naming and representation
of a tripeptide are shown. l. Determination of am_ino acid cornPGlltle.t
in a protein : The protein or polypeptide fs
Dimensions of a peptide chain : ~he comp letely hydrolysed .to. liberate• the amino
dimensions of a fully extended polypeptide acids which are quan_t,tat,ve 1y . estrrnated. The
chain are depicted in Fig.4.7. The two adjacent hydrolysis may be earned out either by acid or
a-carbon atoms are placed at a distance of 0.3 6
alkali treatment or by enzyme hydrolYsfs,
nm. The interatomic distances and bo nd angles
are also shown in this figure. Treatment with enzymes, however results In
smaller peptides rather than amino acids.
Determination of primary structure Pronase is a mixture of non-specjffc
The primary structure comprises the identi- proteolytic enzymes that causes complete
hydrolysis of proteins.
fication of constituent amino acids with regard
to their quality, quantity and sequence in a Separation and estimation of amino acida :
protein structure. A pure sample of a protein or The mixture of amino acids liberated by protein
a polypeptide is essential for the determination hydrolysis can be determined by chromato-
of primary structure which involves 3 stages :
graphic techniques. The reader must refer
1. Determination of amino acid composition. Chapter 41 for the separation and quantitative
determination of amino acids. Knowledge on
! --•-1 PAIHa.. AND AMINO ACIDI

~Q .....,-..,e
2

~,__ _ Polpplde
( N b.lllllllt AA)
R1

02N-Q~~ 6H-COO- ~-tH

-
02
H
Dlnllrophent9 (DNP)-
amlno lCld
0- 'i N

PhlnyllhlohydllllDln (P'TH)-
_,,..~H

l
Identified by
chromatography
amlno acid

l
lclentltled by dN OfflllfiDgfllpt

,.,,.,.., fNOflffl (1-fluoro 2 , ~ ] : _ i t l d I

dJ~Jil'!fbu,lamll!f:~!!l~!fJ- 1· Ill
•a:.-. ~--- -·

primary structure of proteins will be incomplete (c) Breakdown ofpolyptptill a inlo

without a thorough understanding of chromato- fra,nents : Polypeptides are deJ,aded
graphy. into smaller peptides by erµymatic or
chemical methods.
2. Degridation of protein into smaller frag-
ments : Protein is a large molecule which is Enzymatic cleavage : The proteolytic enzymes
sometimes composed of individual polypeptide such as trypsin, chymotrypsin, JlePlift and
chains. Separation of polypeptides is essential elastase exhibit specificity in cle;wiflt •
before degradation. peptide bonds (lefer fis.1.7). Among theta
enzymes, trypsin is most commonly ~ ll
(a) liberation of polypeptides : Treatment
hydrolyses the peptide bonds containing t,,.,.
with urea or guanidine hydrochloride
or arginine on the carbonyl (-C=O) side of
disrupts the non-covalent bonds and peptide linkage.
dissociates the protein into polypeptide
units. For cleaving the disulfide linkages Chemical cleavage : Cyanogen brom;<Je
between the polypeptide units, treatment (CNBr) is commonly used to split p o l ~
with performic acid is necessary. into smaller fragments. CNBr specifically tphts
peptide bonds, the carbonyl side d which ~
(b) Number of polypeptides : The number of
contributed by the amino acid methionine.
polypeptide chains can be identified by
treatment of protein with dan,yl chloride. 3. Determination of . . . acil •IINl.11111•I. ..
It specifically binds with N-terminal amino The polypeptides or their smaller
acids to form dansyl polypeptides which conveniently utilized for the
on hydrolysis yield N-terminal dansyl sequence of amino acids. Th,ls II
amino acid. The number of dansyl amino wise manner to finaU, 1,ul,a
acids produced is equal to the number of amino acids in a
polypeptide chains in a protein. employed for
ptlD•
 e1CJLiMctv11t:t r,::ry

I peptide chain. By analys 1"8

56 acids rn a po Ynce of oNA that codes fior
arnrn 0uc1eot1 de 5equebl to translate the nucleotide
ed 1 nuoro
2, the n ss1 eno acid sequence. This
Sanprs mgfflt: Sang r us nsulin rein, rt ,s P0
pro . to arnr
fails to ident1.fy the disulfide
1
1nitrooMl't'"'<' FONB) to determine~ \ ith equence ,n
u FONB pectficall} bind I s however, . h .
,echn1que, that occur rn t e amrno acids
nninal mmo a 1d to form a dinltroph~:rs bOnds and ch~n~es ynthesized (post-translational
NP dem t1ve of peptide This on hydro
nd r {ter the protein ,s s
\ ds DNP..ammo 1d (N terminalJ a ~ee a . )
mo ,ds from the rest of the peptide chain. rnodificauons .
DNP mrno acid can be 1denttfied b} chrornato· y STRUCTURE Of PROTEIN
fl
ph 5ecoMPA tion of polypeptide chain by
!> n e s reagent has l1m1ted use since the
.Th.e conr ffolding
orrna is referred to as secondary
pept de chain 1s hydroly~ed to amino acids. twisting O h rnino acids are located close to
Edman's reagent: Phenyl isothiocyanate is structure. T e• a their sequence. Two types of
the Edman s reagent. It reacts with the N- each other rn ctures a.-he/Jx A heet, are
. an d ..,-.s
t rmmal ammo acid of peptide to torm a phenyl secon dary strU '
th ocarbamvl derivative. On treatment with mild mainly identified.
c d phen I thiohydantoin (PTH)-amino acid, a Indian scientist Ramachandran made a
cycl c compound is liberated. This can be significant contribution in understanding the
1dentrfied by chromatography (fig.4.lfl. spatial arrangement of polypeptide chains.
Edman s reagent has an advantage since a
peptide can be sequentially degraded liberating
N-termrnal amino acids one after another which a-Helix
can be identified. This 1s due to the fact that the a-Helix is the most common spiral structure
peptide as a whole is not hydrolysed but only of protein. It has a rigid arrangement of
eases PTH-ammo acid. polypeptide chain. a-Helical structure was
Sequenator : This is an automatic machine to proposed by Pauling and Corey (1951) which is
determme the amino acid sequence in a regarded as one of the milestones in the
polypeptide wrth around l 00 residues). ft is biochemistry research. The salient features of
based on the prmcrple of Edman's degradation
a-helix (Fig.4.9) are given below
described above}. Amino acids are determined
equent,ally from N-terminal end. The PTH- 1. The a-helix is a tightly packed coiled
ammo acid liberated is identified by high- structure with amino acid side chains extending
performance lrqurd chromatography (HPLC). outward from the central axis.
Sequenator takes about 2 hours to determine
2· The a-helix is t b1·r
each ammo acid. hydrogen bonding It . ~ a ,zed by extensive
Overlapping peptides attached to peptid~ ~s ormed between H atom
peptide C. The h d , and O atom attached to
In the determination of primary structur
protein, several methods (enzymatic or chem~ of weak but collec/ ~ogen bonds are individually
stabilize the hel' ve y, they are strong enough to
iare simultaneously employed · Th'1s results in cal)
th IX.
ormatlon of overlapping peptides Th' . e
the spec1frc action of different age~ts ~~ ,~_due to last3. ·All the pept,'d e bonds
sites m the polypeptide • Over 1app,ng • ifferent
. h d 111 a polypeptid h ~xcept the first and
are very useful rn determining th ~pt,des Y rogen bonding. e c am, participate in
sequence. e amino acid
_4. Each turn
acids and of ex-helix c .
Reverse. sequencing technque I spa . travels a d' ontams 3 .6 amino
cing of each . rstance of O54 The
It 1s the genetic material ( h .
. h I. amino acid . . nm.
c em,cally D
wh1c u trmately determines the NA)
sequence of spontan
s. a-Heli . rs 0.15 nm
x rs a stabl .
eously With th le conformation formed
e owest energy.
n, Jtaa 4 AND AMNJ A0D8 57

Glu) or basic (L"YJ, Arg. His) am,no adch ao

l111etfete with a-helax 5truetufe.

J}-Pleat ed ~

This ts the second type of 5trudure (hence J'

after a) propmed by Pauling and Corey.
~Pleated sheets (or s,mply Jkheels) are
composed of two or more sea-nents of fully
extended peptide chains {RJ.4.1 ... In the
Jkheets, the hydrogen bonds are formed
between the neighbouring segments of
polypept ide chain(s).

Paralle l and anti-pa rallel iJ.ah ■ ■ts

The polypeptide chains in the ~ may

be arranged either in parallel (the same
direction) or anti-parallel (opposite direction).
This is illustrated in fii-4.10.
~Pleated sheet may be formed either by
separate polypept ide chains (ti-bonds are
interchain) or a single polypept ide chain folding
back on to itself {H-bonds are intrachain).

0 H
II I
{A)

II I II
Q H. j O H-8and
H?i bat 111..ct11n
~
N~
0 tt
~ I
0 H
Ff9. U: Dia,gtammlllic fllP(8NIUIJon of MIOOldlly
6lnJclure ol p,_,,,__. lf/1t hllndlld a-htllbc {8) N-Teminal - - - - - _
••-••-. -. _.., .•t
~ C-l&i.....

,..,....,. ...
t• -h:fcat -C-A
,.,,,,,.dotad,,, ..
lf0UPII~ .,,,,,,, .:Ida;

,.,... , ..,,,,,,..,,,,
.,.~bt inds;
• • • • • • ••••••• I .

(C) N-Tenninal - - - - - - - •
t
C.11••
C-Tennilal .__ _ _ _ _ _ _ N • • • •
6. The right handed a-helix is more stable
than left handed helix (a right handed helix turns
in the direction that the fingers of right hand curl
when its thumb points in the direction the helix
rises).
(C)M1(cl I,,..
bofldt b1l1.. ,~U

7. Certain amino acids (particularly proline)

disrupt the a-helix. Large number of acidic (Asp,
 be identical or
1 maY d l'
58 5 whiC \s are terme has ~d•~o~er,
,ptit' e I protei cture. T e rn 1v1dua1
pol> ',
lfl~ •
',~ ted, sue i.,tcrnarY
(jUu nown as monomer,I
stkru
u, d µosst'ss i,ains are dimer consits of t..-o
,,nlyµt'Pt/de \ubunits. A mer has four.
pv ners or 1'le a tetra
protoi tides wh structure : The
µolypeP • quaternahryld together by non.
d5 m • are e b
ubun1ts hydrogen onds,

~
9011 .

--
111 merrc s name Iy . b d
on° bonds . and ionic on s.
convalent . interactions .
d ophobrc ems : These
- ------==:;;.; protein
hy r O f oligom eric prof I .
ntshon ofs tance t role in the regu at,on
t:,g .f..1I : D grammstlc mp~ed sheet (blue). _
18 1rnpor . nif,can
. . .
cants.In 'Ilg a-he 11. and fJ-p/88 . s play a s1g 1/ular function.
protein . m and ce
proteins of metabolrs . ric proteins : Hemo-
Occurrence of ..,-s A ·heets: Many
h the a-he 1·,x Of ohgome I I
oontarn 13-pleate s d heets . As sue ,
d in the sam e Examples
. aspartate transcarbomy ase, actate
globrn,
and 13-sheet are commonly foun the globular
dehydrogenase.
protem .structure {fig.4h11). eInstructure.
protems, (3-sheets form t e cor 'd
responsible for
tures . Bes, es Son ds
Other f) pes of secondary st~ucd ab~ve, the protein structure
the ex-and JJ-struetures _d_escnbl:ss organised
.A bends and nonrepet1t,ve ( d. Protein structure is . stabi Iized by two types of
structures)
ir secondary structures are also foun ,n bonds-cova en I t and non-covalent.
proteins.
1 CovaIent bonds ·. The . disulfide
peptide and
TERTIARY STRUCTURE OF PROTEIN bonds are the strong bonds in protein structu~e.
The three-dimensional arrangement . of The formation of peptide bond and its
protein structure is referred to as tert,~ry chracteristics have been described.
structure. It is a compact structure with
hydrophobic side chains held interior while the Disulfide bonds: A disulfide bond (-S-S) is
hydrophilic groups are on the surface of the formed by the sulfhydryl groups (- SH) of two
protein molecule. This type of arrangement cysteine residues, to produce cystine
ensures stability of the molecule. (Fig.4.12A) . The disulfide bonds may be formed
Bonds of tertiary structure : Besides the in a single polypeptide chain or between
hydrogen bonds, disulfide bonds (-S- S), ionic different polypeptides. These bonds contribute to
interactions (electrostatic bonds) and the structural conformation and stability of
hydrophobic interactions also contribute to the proteins.
tertiary structure of proteins. 2
Domains : The term domain is used t 1 · Non-covalent bonds : There are, mainly,
represent the basic units of protein st t 0 our types of non-covalent bonds.
rue ure
. A JlO/ypeplide With
(tertiary) and function. (a) Hydrogen b d
amino acids normally consists of tw 200 are fornied bon hs : . The hydrogen bonds
domains. 0 or more
betWeen th Y s aring
. of hydrogen atoms
nitrogen and carbonyl
oxyg en of e d"ffi
QUATERNARY STRUCTURE OF pb I
(Fig,4, 128) E erent peptide bonds
A great majority of the proteins 110fEIN
but collect/ve~ch hydrogen bond is weak
of single polypeptide chains are con,Posed th
proteins, however, consist a(· t5ome Of the nuniber of h Y ey are strong. A large
contribute IO Yirogen bonds significantly
wo or more
(bJ fiyd t e Protein structure.
rophobic bo
nds : The non-polar side
chains of ne
lJtraJ amino acids tend to be
°"lllllli • IN AND AMINO AC101

~NH,:00....vv"\ acidic ,mtno acldt wNh

charpd IIOUPf (1.1. •NHf> 4 ...
amino acids (llf,4-flOJ,
p,..,,
w.l t ,._ , n. , • •

~.h:~
(d) VII . ,
non•CO\llln MOCfadoftl ~••••n
electrlally neucral molecull,. n., •
formed by 1M tlec:bUlltk if•ac:llonl
due to permanent or induced dlpcM.
(I)
~ i- r,
. .
-~ ~ lxam pl•• of prot ein 9ffllCIUN'
Structure ol .... .... ... : ,.,,. ,. COfllillt
H f\ 6 of two polypeptide chains, A and B (14f .U1
~~ -6 H- 8/ "v 'vV The A chain has 1lyclne at the N IBftDinal ...
and aspara1ine at the C-terminal end. The I
(C) ~NH-yH-CO~ chain has phenylalanine and alanine al the
N- and C-termlnal ends, respectively. Oriafnaltf,
HC CH 3
insulin is synthesized as a sinpe ..,.,....le
";lie
preproinsulin which 1,,.det.,es µ
processing to give proinsalln and 811111, ,,,,,,,._
The structural aspects of henqlollin and
collagen are respectively giwn iii C 3 f 1 •
and 22.

Methods to determine
(D) protein structure
For the determination of secondlly and
tertiary protein ~re s, X-ray ~
is most commonly used. Nuclear mat9lelk
resonance (NMR) spectra of proteins pKHidls
structural and functional infonnatian • ._

,,.,,,, .. 111 11111

•··~~•ii~IIM1111
"" -'IWlliil •~
atoms and lfOUps present in the p;uWei$

7 s-s]
..;
. .; 1.. _~...;&-?:.-. .
, _ ._...,.._ _.....
I 11 f •l1 •-
S S
closely associated with each other in I
proteln1 (FIJ.4. f ZC,. As such, these are I
not true bonds. The occurrence of
ls ls
hydrophobic forces is ohlerved in 1•
aqueous environment wherein the 7
molecules are forced to stay topther.
(cl ladr I bdk ltondl I These bondl are
formed by Interactions between
nepttvejy cha,pd lf'OUPI (e.a. COO-} al
 . caseln-4.6; Human alburnf
Pepslr1-1.' ~emoslobin-6.7; Lysozyrne-11
.......... for the teolatlon urease-5,0,
ncl .,_.c proteins : Protef
MNI Plll'ffleatton of protein•
~ a l methocf5 are employed to i50l~te
purify ~ n s fnftfafly proteln5 are fradion~
a: 1
8
S. Addle: (£ Lys + £ Arg)/(e Glu + 1
which the rat,° re referred to as basic
greater than ;ins the ratio is less than 1
Pre_.
by uslft8 d ~ t conc:entratlons of ammoni~m For acidic pro ' '
sulfal@ or sodium sulfate Protein fradlon~tion red itatian of proteiM: Proteins e,c
may af,o be carried out by ultracentrifugation. 6.. P Plution due to hydration of
colloidal so NH+ -OH) Proteins
Protein separation 1s achieved by utilizing (-COO-, - 3, . .
~~s 1soelectric focussing, immuno- grou~~ ed b dehydration or neutrahzatiot1
prec1p1tat Y
electruphores15 eon-exchange chromatography, polar groups.
&el frration, high performance liquid ch~ato- ,rec:ipitation at pl: The pro!eins in
ltaphy (HPI.Q etc. The details of these techniques iubie at isoelectnc pH. C
are I.eas(t SOcasein) get eas1·1 y prec1p1
• 'tated
aft! described 1n CJupte,- "'· proteins e.8 · t
the pH is adjusted to P! (~.6 or c
PROPERnEs OF PROTEINS . of curd from milk 1s a .marvel
Format,on .
1 Salulaility : Proteins form colloidal exampIe Of S
low precipitation of milk
.
solutJons rnstead of true solutions in water. This casein· at pl • This occurs due to the lactrc .
rs due to huge size of protein molecules. uced by fermentation of bacteria w
prsod f .
lowers the pH to the pl o casein.
2. Molecular weight: The proteins vary in
the r molecular weights, which, in turn, is Precipitation by sahing out : The process
dependent on the number of amino acid protein precipitation by the additional of
residues. Each amino acid on an average salts such as ammonium wlfate or miurn
contributes to a molecular weight of about 110. is known as salting out. This phenomen
MaJorrty of proteins/polypeptides may be explained on the basis of dehydration of
composed of 40 to 4,000 amino acids with a molecules by salts. This causes increased nrrllMI
molecular weight ranging from 4,000 to protein interaction, resulting in mol
440 000 A few proteins with their molecular aggregation and precipitation.
werghts are listed below :
The amount of salt required for p
lnsuhn-5,700; Myoglobin-1,700; Hemoglobin- precipitation depends on the size (mol
64,450 Serum albumm-69,000. weight) of the protein molecule. In general,
3 Shape : There is a wide variation in the higher is the protein molecular weight, the
protem shape It may be globular (insulin), oval is the salt required for precipitation. Thus,
(albumin) fibrous or elongated (fibrinogen). """"1• are pred,,.ted by luff,.,.,,.,..,
4. ltoelectric pH: lsoelectric pH (pl) as a ammonium sulfate while albumin is preci
property of amino acids has been described. The by lull •turatlon. Salting out procedu
nature of the amino acids (particularly their conveniently used for separating serum al
from globulins.
1on1zable groups) determines the pl of a protein.
The acidic ammo acids (Asp, Glu) and basic The addition of small quantities of
amino acids (His, lys, Arg) strongly influence the salts Increases
the solubility of proteins.
pl At ,soelectr,c pH, the proteins exist as
n,,..,.,_ Pr0Cess called as _,.,, In is due ID
or dlpolar Ion,, They are electrically diminished Prot •
neutral (do not mlsrate In lhe electric fletd) with concentratton. ein-protefn Interaction at
minimum so,ublllty, maximum PNC:ipltablllty
and' leut bidferlng apacJty 11w IIOllectrJc
pH(pl) tor ,o,ne p,del111 a,e alven "-
 61
Ch■ptw 4 . PFl.Jlt:1-..:::; At#:J AMNJ AOOS

bemg PoS1t1vely charged \'Ii hen added ro protean b1uret test 1s not de.arty known It 1s believed
solution (negauvel) charged) m alkahne medium 1hat the colour ,s due to the ronnar,on of a
results in prec1p1tate formatron copper UHH diut« I compk:x as shown below

Prttipifation by anionic or albJoid reagents : 0 0

Proteins can be prec1p1tated by tnchloroaceuc NII C
/c NII
acid sulphosaficyl,c ac,d phosphotungst1c acid
p1cnc acid, tanmc acid phosphomol).fxf1c acid
etc. B) the add1t1on of these acids, the
proteins existing ac: cations are prec1p1tated b}
the anionic form of acidc: to produce protein-
\ 0
NH
'ct,2·

Ml C
0
"/
NH

sulphosafiq late protein-tungstate, protein-

picrate etc.
Precipitation by organic solvents : Organic
solvents such a, alcohol are good protein
precipitating agents. The} dehydrate the protein
molecule by removing the \\ ater envelope and
cause precipitation.
7. Colour reactions of proteins: The proteins
gh e several colour reactions which are often
useful to identify the nature of the amino acids
present in them.
The presence of magnesium and ammonium
ions interfere in the biuret test. This can be
m ercome b) using excess alkali.
The colour reactions given by proteins due to
the presence of specific amino acids are given in
fable 4.3. These reactions are often useful to
know the presence or absence of the said amino
acids in the given protein.
DENA TURA TION

Biuret reaction : Biuret is a compound formed The phenomenon of disorp nwtio n of ru~
b}' heating urea to l 80°C. protein structure is known as denaturation.
Denaturation results in the loss of secondary,
NH2 tertiary and quaternary structure of proteins. This
C=O
I
~H2 involves a change in physical, chemical and
I C-O biological properties of protein molecules.
NH2~aocc I -
+ NH
I
NH2 C=O
I I TAil£ 4.3 Colas re.idimls of proteills/MlillO «MS
y=O NH2
NH2 ReM:tion Sp«i(i c JfOUP or._, Mid
2 Moles of urea Bult
1. Binet reactioo Two peptide lnlgas
When biuret is treated with dilute copper 2. Nilh)ml reaction a-Amilo acids
sulfate in alkaline medium, a purple colour is 3. Xanthopnuic Benzene mg d nnllic
obtained. This is the basis of biuret test widely reaction Mn> acids (Phi, Tyr, T,p)
used for identification of proteins and peptides.
4. Milorls reaction Pheuoic ~ (Tyr)
Biuret test is answered by compounds ---- ---- ---- ··-lndole mg (Trp)
containing two or more CO-N H groups i.e.,
s. Hopki1s-Cole raaction
• reaction
peptide bonds. All proteins and peptides 6. Ir

possessing at least two peptide linkages i.e., 7. Nilroprusaide raadion

tripeptides (with 3 amino acids) give positive
biuret test. Histidine is the only amino acid that 8. Solfurtast
answers biuret test. The principle of biuret test is 9. Pauty'stast
conveniently used to detect the presence of 10. Foin-Coiallllu's 1111
proteins in biological fluids. The mechanism of
 kl 1~ i\UIM protein
bonds to t'n \ nw§ ( on ' •·tooked ,___.
d th,,,,,(ort' .., •~
dt'n itur,,Hon '"' f
lpl'Olt'ltl) is nmrt' t \slh dlRl'stN
' 8 Ot-n ltur,,tion Is usu,,lly irrf>verslble F•
k., t~m•t
~ ' ~•lt•t C"An bt• prt'P 1rt1d from an 9II
lbl
ts t: tlN,, nlt't ll~ mshlnl't" om..
pro~in ~,lbumm) but tht' rt'Vt:'rMI I~ not poss •
<>. C.u~tul cfenaturtltion 1s sometimes rever-
a. teristics of ctenaturation .,s
sibh... (t...no\\ n renatur•tion). Ht'moelobln
unclerg0t.'s cfen,,tm,,tion m the pr('sence of
•~"'-•' ~ ture Of protem IS sttlicylate By rt'mov,,I of s,,li<.:ylate, hemoglobtn
1s ren,,tured.
strulctllre ot :1 protean with
ntact I e peptide
Io. Den,1tured protein c<umot be crystallized
Coagulation: The term 'coagulum' refers to a
ts 1cal activity. semi-solid viscous precipitate of protein
Irreversible denaturation results in coagulation
Denature,a nirntem becomes insoluble in
Coagulation 1s optimum and requires lo\9811
t was ongmalfy soluble.
temperature at 1soelectric pH. Albumins and
of denatured protean globulins (to a lesser extent) are coagulable
while ts surtace tension proteins. HHt coa,uution Int Is ~
uted to defffl th. presence of albumin In
assocYted with lncf6se in Flocculation : It is the process of
ioniz.able and groups of Protein This
lfhydryf Pfeetpitation at 1soelectr1c pH. The precipitaW
due to loss hydropn •nd disulfide bonds.
referred to as flocculum. Casein (millc p,011ht
7 Denatured p,olein • more easdy di&ested. can be easily Precipitated when adju11...t'9111
Thts as due to incrNSed exposure of peptide ~~ectric PH (4.6) by dtlute acetic
OCC:ulatlon is tevenible On applicat
Ch■11Ns 4 PRCU&a NG AMINO ACID8

7 Glll fl pUJI L Nrad J f 1,

I Df psal I s. . - ~ Iii[. . .a.
O(lla bufn
CLA naFI CAT ION OF IIIIO JIIN 8
Proteins are classified in severa l ways Three 9. 11 I ,•11 llf•' for..........,
...a.
Yitn

~,or types of clasHytng proteans bned on lheir IL ..,_ . of■ rrNl1dlM ~nss ll •
f u ~ chemical nature and solubility CNffllcal ..... .. and ... 1 . ,
propertaes and nutrit ional intpCN tance are
,lN!iBWe
__..
. - 1111,a•r...•
,
discussed hen!. This is a more c:oo.p
classificalion al p,Ole int. • • ...., OIi . .
, lllalpe _,
A. Func tloew l CINaltlcatio.t of prot eins amino acid camp o5ilio n, . . . . _
they perfo.m, proteins solubility p.upa ties. ~c1e· 111 ae kJ #\
Based on the functions
classified inll> 3 major poups
are classified into the following groups. (with
examples) 1. s· i I pi 1 • : They ae GW4 i E1 fll
_,, • at:itl R!5idlres
1. StructunP pculeiaw : Keratin of hair and
nails, collagen of bone. 2. Cc j J I I fl I • : B! . k 6e zl ■
acids, these proteins UNltlin a ,at; 1 •
2. Enzymes °' catalytic pnlteiaw : Hexokinase, moiet y known as F d 1 ; O •
pepsin. .
conjugating group
3. Transpo.1 proteins: Hemoglobin, serum 3. De.Z-» p:e• • : These ae th... cha e al
albumin. or deg,aded prodllds of simple and cu i a I
4. Hormonal pro1ein1: Insulin, growth proteins.
hormone. The above three d er ae ._,_
5. Contractile proteins : Actin, myosin. subdivided into diffelent p,up s. The • LWf
6. Storage proteins : Ovalbumin, glutelin. of prolei n classification is gi"9I in the Tallt U.

Proteta • ,,_ , . . . , . , . . Olpl k •tr11th o1

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ffw . . . JullflflfillMI t111111Mf11 -

"21 a
clofflns ;.to,,, tmmw

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Holl of ,,_ o
..•.•,,
~ ~ J O , ,,_