Chng 2: PROTEIN

Axit amin Cng th c chung Phn lo i aa Tnh ch t l ha Peptide C u t o v tnh ch t M t s peptide quan tr ng Protein nh ngha, ch c nng protein Lin k t ha h c trong phn t protein C u trc phn t protein Phn lo i protein S bi n tnh protein

- Axit amin
Ch c nng
n v c u trc c a protein Ch t d n truy n th n kinh (neurotransmitter)

C u trc chung
Nhm carboxylic Nhm Amine Nhm ngo i R T i v tr C

C 20 axit amin tham gia vo c u trc protein

-axit amin
Phn lo i axit amin d a vo:
Tnh phn c c (polarity) Nhm ch c nng (functional groups) Tnh axit-baz-trung tnh

Tnh ch t c a axit amin

Ha h c l p th
Axit amin l ch t ho t ng quang h c, c kh nng quay m t ph ng nh sng H u h t axit amin c a protid thu c dy L

Tnh ch t c a axit amin

Tnh l ng tnh
Trong mi tr ng n c, axit amin phn li hon ton Tc d ng v i axit v baz m nh t o thnh mu i i n tch axit amin ph thu c vo i n tch c a mi tr ng i m ng i n pI l i m pH nh t nh m t i t (-) b ng dt (+) Axit amin khng t n t i d ng t do m d i d ng mu i n i
H H2N C R COOH

H NH3
+

C R

+ COOH +H NH3

H C R

-+HO COO H2N

H C R COO
-

H2O

Dang cation pH<7

Ion lng cc pH=7

Dang anion pH>7

B ng 1: Gi tr pKa v pI c a cc axit amin

Amino acid Glycine Alanine Leucine Isoleucine Serine Threonine Asparagine Tryptophan Valine Glutamine Methionine Phenylalanine Proline Aspartic acid Glutamic acid Histidine Cysteine Tyrosine Lysine Arginine
pK1 -COOH pK1 -NH3 pK1 R group pI

2.34 2.34 2.36 2.32 2.21 2.63 2.02 2.38 2.29 2.17 2.28 2.58 2.00 2.09 2.19 1.82 1.71 2.20 2.18 2.17

9.6 9.69 9.60 9.76 9.15 10.43 8.80 9.39 9.72 9.13 9.21 9.24 10.60 9.82 9.67 9.17 10.78 9.11 8.95 9.04 3.86 4.25 6.6 8.33 10.07 10.53 12.48

6.07 6.07 6.64 6.64 5.68 6.16 5.41 5.88 6.00 5.70 5.74 5.91 6.30 2.80 3.22 7.59 5.02 5.66 9.74 10.76

Tnh ch t c a axit amin

Tnh ch t l h c
Tnh tan trong dung mi phn c c: n c, ethanol 75-80%, Khng h p thu b c sng Vis ho c UV c > 240nm, ngo i tr cc aa vng nh tryptophan, tyrosin, histidin, phenylalanin Th ng c v ng t ki u ng

Tnh ch t ha h c
Ph n ng do nhm -carboxyl v -amin Ph n ng c a nhm -amin Ph n ng c a nhm -carboxyl

Ph n ng do nhm -carboxyl v -amin

Ph n ng Ninhydrin
Nhi t cao ~100oC NH3, cc peptide ph n ng ch m hn so v i axit amin Prolin, hydroprolin t o ph c mu vng v i Ninhydrin Pht hi n -axit amin (phng php s c k gi y)

Ph n ng do nhm -carboxyl v -amin (tt)

Ph n ng ngng t t o lin k t peptide
-amin c a aa ny v i carboxyl c a aa khc T o chu i peptide

Ph n ng c a nhm -amin
Ph n ng Sorensen
V i formaldehyde
R H C N H2 H R R
,

+O

N=CH-R

COOH

H2 O

Foocmaldehyt

COOH

Kha nhm -NH2 c a axit amin nh l ng COOH b ng phng php chu n Suy ra hm l ng c a axit amin

Phng php xc nh th t amino acid

Phng php Sanger
Pht hi n 1953 Xc nh aa u N b ng 1-fluoro-2,4-dinitrobenzen (FDNB) D t s i polypeptide

Ph n ng c a nhm -amin
Ph n ng v i axit nit, HNO2
nh l ng axit amin theo l ng kh N2 t o thnh

R HC NH2

HNO2

HC

OH

N2

+ H2 O

COOH

COOH

Ph n ng oxy ha kh nhm amin t o thnh xetoaxit

COOH H C R NH2

COOH

+ 1/2 O2

CO R

NH3

Ph n ng c a nhm -carboxyl
Ph n ng kh nhm -COOH

R H C N H2 -CO2 H COOH

R C N H2

CO2

COOH

Peptide
C ut o
Phn t c u thnh t 2 hay nhi u axit amin lin k t v i nhau b ng lin k t petide Vd: Insulin: 51 aa; Lysozyme: 129 aa; trypsinogen: 231 aa

Peptide
Phn lo i
Dipeptide: 2 aa Tripeptide: 3 aa Oligopeptide: < 10 aa Polypeptide: 10 aa

Peptide
Tnh ch t
Ph n ng Biure

Protein Peptide ( 3 aa)

Cu2+ OH-

Peptide
M t s peptide quan tr ng
Carnosine
Dipeptide: beta-alanin-L-histidine M c v no Anti-oxidant, anti-glycation, chelator of metal cation (Cu2+)

Glutathion
Tripeptide c cng th c: -glutamyl-cysteinyl-glycin Nhm ho t ng: nhm SH c a cystein Tham gia vo h oxy ha-kh C nhi u gan, th n, l lch

Protein
nh ngha
M t trong nh ng i phn t sinh h c khng th thay th c a c th s ng Protein c th g m m t hay nhi u chu i polypeptide s p x p theo m t tr t t xc nh. Chu i polypeptide l m t polymer v i n v c u trc l cc phn t axit amin

Protein
Ch c nng
Xc tc: enzyme V n t i: hemoglobin Chuy n ng
S co c: chuy n ng tr t ln nhau c a 2 l o s i protein: s i tomiozin, s i m nh-actin,troponiozin v troponin

B o v : khng th , interferon Truy n xung th n kinh: rodopxin mng m t i u ha: truy n thng tin, trao i ch t, bi u hi n gen Ki n t o ch ng c h c: sclerotin trong l p v ngoi c a cn trng D tr dinh d ng: cung c p aa cho s pht tri n c a phi

Protein
Tnh ch t
Phn t l ng l n Ch t i n li l ng tnh (nhm ngo i R) pI l pH m t i t ng s t(-) = t(+), protein trung ha v i n
pH<pI, protein tch i n dng pH<pI, protein tch i n m

Tinh ch t dung d ch keo protein

Trong n c, protein bi hydrat ha, v hydrat c b dy 3 d ch keo Pp th m tch tinh s ch protein b n dung d ch keo phu thu c:
S tch i n cng d u c a protein (pH pI) L p v hydrat bao quanh phn t protein

dung

(?) K t t a protein

Kh i lng phn t v pI c a m t s protein

Protein Xitocrom C Ribonuclease Tripxin Pepsin Hemoglobin Albumin huy t thanh M, dalton 11 600 12 700 24 000 36 000 64 500 69 000 pI 10.6 7.8 10.5 1.0 6.8 4.9

Cc lo i lin k t ha h c trong phn t protid Lin k t peptide (peptide bonds)

Lin k t c ng ha tr quan tr ng nh t, G = -300 ~ -400 kJ/mol T o khung cho chu i protein

Cc lo i lin k t ha h c trong phn t protid (tt)

Lin k t disulfit (disulfit bonds)
Lin k t cng ha tr quan trong th 2, gi vai tr t o hnh v ch c nng cho phn t protein Lin k t cho hnh thnh ngay c gi a cc ph n khc nhau c a cng 1 chu i polypeptide ho c 2 chu i khc nhau Lin k t gi a 2 nhm ngo i aa cysteine qua nhm ho t ng sulphydryl SH t o thnh nhm ngo i c a aa cystine

Cc lo i lin k t ha h c trong phn t protid (tt)

Tng tc ion (ionic interaction or salt bridge)
Tng tc hnh thnh gi a nh ng nhm mang i n tch tri d u khi chng g n nhau tng i, ~0.3nm Vd: tng tc ion gi a nhm COO-, nhm ngo i aa glutamic acid v NH3+, nhm ngo i c a aa lysine
G = -4kJ/mol

Cc lo i lin k t ha h c trong phn t protid (tt)

Lin k t hydro (hydrogen bonds)
Lin k t hnh thnh gi a >N H v >CO ; G = -3kJ/mol Lin k t n, c ngha l n trong i phn t
T o c u hnh khng gian v hoat tnh sinh h c c trng

Lin k t y u, hnh thnh v b gy m t cch d dng nhi t bnh th ng

Ch t trng h p (protein v a. nucleic) c c u trc linh ho t, thch nghi trong i u ki n c th Tnh b o t n v bi n d trong di truy n

Cc lo i lin k t ha h c trong phn t protid (tt)

L c Van der Waal
L c h p d n gi a cc nguyn t Xu h ng ko cc nguyn t n g n nhau hn

Cc lo i lin k t ha h c trong phn t protid (tt)

Tng tc k n c
Th hi n xu h ng lin h p c a nh ng nhm k n c v i nhau Lo i n c ra kh i phn t protein tng entropy (S) Nhm ngo i k n c

C u trc phn t protid

C u trc b c 1: trnh t s p x p c a cc aa trn m ch polypeptide nh m i lin k t peptide
Gi s : 1 chu i polypeptide c 100 aa c th c 20100 chu i polypeptide B t c m t s thay i no trong c u trc b c 1 gy ra m t s t bi n, b nh t t
HbA ng i bnh th ng: HisVal-Leu-Tre-Pro-Glu-Glu-Ly- HbS ng i b b nh hnh c u l i li m: thay th Glu(6) b ng Val(6)

C u trc phn t protid (tt)

C u trc b c 2
S s p x p thch h p trong khng gian c a m t chu i polypeptide do Ca c kh nng quay t do xung quanh tr c c t o thnh t cc lin k t c ng ha tr i u ki n nh t nh, hnh th t nhin c a phn t protein t n t i sao cho nng lng t do (G) c a h th ng t c c ti u

C u trc b c 2 th ng g p
C u trc xo n c: xo n , , , 310
Xo n : tng t l xo 3.6 aa/vng ~ 18 aa/5vng xo n Gc xo n: 26o, b c xo n: 0.54nm Xo n ph i/ xo n ph i Lin k t hydro t i a: nhm NH c a lk peptide ny v i nhm CO c a lk peptide th 3 k v i n C u trc c tr t t v r t b n v ng Xo n , th c 4.4 v 5.2 g c axit amin trong m t vng Xo n 310 l d ng xo n a v i 3 g c axit amin/vng

C u trc b c 2 th ng g p (tt)
C u trc g p n p
Hnh ch chi Xo n g p n p C 2 lo i g p n p:
Song song i song song

Lin k t hydro Gc g p: 130o, n p g p: 3.47

C u trc b c 2 th ng g p (tt)
C u trc xo n collagen

C u trc cu n ng u nhin
C u trc khng xc nh, khng c m t ph ng v tr c i x ng Nhm ngo i R mang i n tch ho c b n ng khng gian

C u trc phn t protid (tt)

C u trc b c 3
S phn b trong khng gian c a m ch polypeptide c ch a cc o n xo n -helix v -sheet Nh ng lin k t n i phn t lm cho c u trc b c 3 b n v ng: tng tc ion- c u mu i; lin k t hydro; lin k t disulfit, Qui nh hnh d ng bn ngoi v ho t tnh sinh h c c a protein VD: lysozyme, ribonuclease, myoglobin
Myoglobin Lysozyme Ribonuclease

C u trc phn t protein (tt)

C u trc b c 4
C u trc b c 4 l s lin h p c a cc ti u th t o thnh phn t l n Lin k t hydro, tng tc ion, tng tc k n c ngha quy t nh i v i tnh ch t sinh h c, ho t tnh v s c hi u c a protein

Insulin

Phn lo i
Hnh d ng
D ng s i: hnh thon di, khng tan, b n trong pH
Protein c u trc Collagen
C u trc xo n collagen (collagen helix), xo n tri Tropocolagen: 3 chu i polypeptide, 1000aa/chu i Thnh ph n chnh c a gn, xng, da, rng v m x n

Silk
C u trc -sheet

Keratin
Protein thnh phn quan tr ng trong tc, t, mng vu t, lng v C u trc xo n hay m ng

Phn lo i (tt)
Hnh d ng
D ng hnh c u: d tan trong n c, nh y v i pH
Protein ch c nng: enzyme, hormon, khng th , Protein cu n l i, s p x p trong khng gian, qui d nh tnh ch t sinh h c c a n

Phn lo i protid
Protid n gi n: ch c u t o t cc axit amin, protein
Albumin
tan trong n c, khng tan trong (NH4)2SO4 c nhi u trong huy t thanh, tr ng, la m, s a,

Globulin
Khng tan trong n c, tan trong d ch mu i long C nhi u trong mu, s a, lng tr ng, m c,

Prolamin
Khng tan n c, c n tuy t i, d ch mu i, tan trong c n 70-80%

Glutenin
Tan trong dd NaOH 2% Protein c a la m

Phn lo i protid
Protid ph c t p (proteic)
C u t o g m ph n protein (apoprotein) v nhm ngo i nh lipid, gluxit v
Lipoprotein = protein + lipid Photphoprotein = protein + H3PO4 qua lin k t ester Glucoprotein = protein + gluxit Nucleoprotein = protein + acid nucleic

S bi n tnh protein
S thay i c u trc 2,3,4 c a phn t protein m khng thay i c u trc b c 1 c a n Tc nhn gy bi n tnh
V t l: tia c c tm, sng siu m, khu y c h c Ha h c: axit, ki m m nh, mu i kim lo i n ng, tanin

Tnh ch t protein b bi n tnh

ha tan gi m, kh nng gi n c gi m M t ho t tnh sinh h c Tng nhay v i s t n cng c a protease Tng nh t n i t i M t kh nng k t tinh

C 2 lo i bi n tnh:
Bi n tnh thu n ngh ch: l bi n tnh m khi lo i b tc nhn gy bi n tnh, protein s tr l i trang thi ban u Bi n tnh b t thu n ngh ch