47

Chng 3

Protein
Protein l hp cht hu c c ngha quan trng bc nht trong c th sng, v mt s lng, protein chim khng di 50% trng lng kh ca t bo; v thnh phn cu trc, protein c to thnh ch yu t cc amino acid vn c ni vi nhau bng lin kt peptide. Cho n nay ngi ta thu c nhiu loi protein dng tinh th v t lu cng nghin cu k thnh phn cc nguyn t ho hc v pht hin c rng thng thng trong cu trc ca protein gm bn nguyn t chnh l C, H, O, N vi t l C 50%, H 7%, O 23% v N 16%. c bit t l N trong protein kh n nh (li dng tnh cht ny nh lng protein theo phng php Kjeldahl bng cch tnh lng N ri nhn vi 6,25). Ngoi ra trong protein cn gp mt s nguyn t khc nh S 0-3% v P, Fe, Zn, Cu... Phn t protein c cu trc, hnh dng v kch thc rt a dng, khi lng phn t (MW) c tnh bng Dalton (1Dalton = 1/1000 kDa, c l kiloDalton) ca cc loi protein thay i trong nhng gii hn rt rng, thng thng t hng trm cho n hng triu. v d: insulin c khi lng phn t bng 5.733; glutamat-dehydrogenase trong gan b c khi lng phn t bng 1.000.000, v.v... T lu ngi ta bit rng protein tham gia mi hot ng sng trong c th sinh vt, t vic tham gia xy dng t bo, m, tham gia hot ng xc tc v nhiu chc nng sinh hc khc. Ngy nay, khi hiu r vai tr to ln ca protein i vi c th sng, ngi ta cng thy r tnh cht duy vt v ngha ca nh ngha thin ti ca Engels P. Sng l phng thc tn ti ca nhng th protein. Vi s pht trin ca khoa hc, vai tr v ngha ca protein i vi s sng cng c khng nh. Cng vi nucleic acid, protein l c s vt cht ca s sng.

3.1. Amino acid

3.1.1 Cu to chung Amino acid l cht hu c m phn t cha t nht mt nhm carboxyl (COOH) v t nht mt nhm amine (NH2), tr proline ch c nhm NH (thc cht l mt imino acid).

48

Trong phn t amino acid u c cc nhm COOH v NH2 gn vi carbon v tr . Hu ht cc amino acid thu nhn c khi thu phn protein u dng L- amino acid. Nh vy cc protein ch khc nhau mch nhnh (thng c k hiu: R).

Hnh: 3.1. Cng thc cu to chung ca cc amino acid

3.1.2. Phn loi amino acid Hin nay ngi ta phn loi amino acid theo nhiu kiu khc nhau, mi kiu phn loi u c ngha v mc ch ring. Tuy nhin, h u da trn cu to ho hc hoc mt s tnh cht ca gc R. V d c ngi chia cc amino acid thnh 2 nhm chnh l nhm mch thng v nhm mch vng. Trong nhm mch thng li tu theo s c mt ca s nhm carboxyl hay s nhm amine m chia ra thnh cc nhm nh, nhm amino acid trung tnh (cha mt nhm COOH v mt nhm NH2); nhm amino acid kim (cha mt nhm COOH v hai nhm NH2); nhm amino acid acid (cha hai nhm COOH v mt nhm NH2). Trong nhm mch vng li chia ra thnh nhm ng vng hay d vng v.v... C ngi li da vo tnh phn cc ca gc R chia cc amino acid thnh 4 nhm: nhm khng phn cc hoc k nc, nhm phn cc nhng khng tch in, nhm tch in dng v nhm tch in m. Tuy nhin, hin nay cch phn loi cc amino acid ang c nhiu ngi s dng nht l da vo gc R ca amino acid v c chia lm 5 nhm: Nhm I. Gm 7 amino acid c R khng phn cc, k nc, l: glycine, alanine, proline, valine, leucine, isoleucine v methionine. (Hnh 3.2)

49

Hnh 3.2. Cng thc cu to ca cc amino acid nhm I

Nhm II. Gm 3 amino acid c gc R cha nhn thm, l phenylalanine, tyrosine v tryptophan (Hnh 3.3.)

Hnh 3.3. Cng thc cu to ca cc amino acid nhm II

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Nhm III. Gm 5 amino acid c gc R phn cc, khng tch in, l serine, theonine, cysteine, aspargine v glutamine (Hnh 3.4)

Hnh: 3.4. Cng thc cu to ca cc amino acid nhm III

Nhm IV. Gm 3 amino acid c R tch in dng, l lysine, histidine v arginine, trong phn t cha nhiu nhm amin (hnh 3.5).

Hnh 3.5. Cng thc cu to cc amino acid nhm IV

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Nhm V. Gm 2 amino acid c gc R tch in m, l aspartate v glutamate, trong phn t cha hai hm carboxyl (hnh 3.6).

Hnh 3.6. Cng thc cu to ca cc amino acid nhm V

3.1.3. Cc amino acid thng gp Cc amino acid thng gp l nhng amino acid thng c mt trong thnh phn ca cc loi protein. Chng c khong 20 loi v c thu nhn khi thu phn protein. Cc loi amino acid ny c tn gi, khi lng phn t v k hiu c trnh by trn bng 3.1. 3.1.4. Cc amino acid khng th thay th Cc amino acid c hnh thnh bng nhiu con ng khc nhau. Nh bit, trong phn t protein c khong 20 loi amino acid, tuy nhin trong c th ngi v ng vt khng tng hp c tt c cc loi m phi a t ngoi vo qua thc n. Nhng amino acid phi a t ngoi vo c gi l cc amino acid khng th thay th. Ngi ta bit c c khong 8-10 loi amino acid khng th thay th bao gm: Met, Val, Leu, Ile, Thr,Phe, Trp, Lys, Arg v His v ngy nay ngi ta cn xem Cys cng l mt amino acid khng th thay th. 3.1.5. Cc amino acid t gp Ngoi cc amino acid thng gp trn, trong phn t protein i khi cn c mt s amino acid khc, l nhng loi t gp. Cc amino acid ny l dn xut ca nhng amino acid thng gp nh: trong phn t collagen c cha 4-hydroxyproline l dn xut ca proline, 5hydroxylysine l dn xut ca lysine v.v...Mt khc, mc d khng c trong cu trc protein, nhng c hng trm loi amino acid khc cng c th tn ti dng t do hoc lin kt vi hp cht khc trong cc m v t

52

bo, chng c th l cht tin thn hay l cc sn phm trung gian ca qu trnh chuyn ho trong c th. Bng 3.1. Cc amino acid thng gp
Tn amino acid Glycine Alanine Proline Valine Leucine Isoleucine Methionine Tyrosine Tryptophan Serine Threonine Cysteine Aspargine Glutamine Lysine Histidine Arginine Aspartate Glutamate Tn amino acid gi theo danh php ho hc -aminoacetic acid -aminopropionic acid -pyrolydilcarboxylic acid -aminoisovaleric acid -aminoisocaproic acid -amino--metylvaleric acid -amino--metylthiobutyric acid -amino-hydroxyphenylpropionic acid -amino--indolylpropionic acid -amino--hydoxypropionic acid -amino--hydroxybutiric acid -amino--thiopropionic acid amid ca aspartate amid ca glutamate , diaminocaproic acid -amino--imidazolpropionic acid -amino--guanidinvaleric acid -aminosuccinic acid -aminoglutarate Tn vit tt Gly Ala Pro Val Leu Ile Met Phe Tyr Trp Ser Thr Cys Asn Gln Lys His Arg Asp Glu K hiu G A P V L I M F Y W S T C B Q K H R D E Khi lng (MW) 75 89 115 117 131 131 149 165 181 204 105 119 121 132 146 146 155 174 133 147

Phenylalanine -amino--phenylpropionic acid

3.1.6. Mt s tnh cht ca amino acid 3.1.6.1. Mu sc v mi v ca amino acid Cc amino acid thng khng mu, nhiu loi c v ngt kiu ng nh glycine, alanine, valine, serine, histidine, tryptophan; mt s loi c v ng nh isoleucine, arginine hoc khng c v nh leucine. Bt

53

ngt hay cn gi l m chnh l mui ca natri vi glutamic acid (monosodium glutamate). 3.1.6.2. Tnh tan ca amino acid Cc amino acid thng d tan trong nc, cc amino acid u kh tan trong alcohol v ether (tr proline v hydroxyproline), chng cng d ho tan trong acid v kim long (tr tyrosine). 3.1.6.3. Biu hin tnh quang hc ca amino acid

Hnh 3.7. ng phn lp th ca alanine

Cc amino acid trong phn t protein u c t nht mt carbon bt i (tr glycine) v th n u c biu hin hot tnh quang hc, ngha l c th lm quay mt phng ca nh sng phn cc sang phi hoc sang tri. Quay phi c k hiu bng du (+), quay tri c k hiu bng du (-). Gc quay c hiu ca amino acid ph thuc vo pH ca mi trng. Tu theo s sp xp trong cu trc phn t ca cc nhm lin kt vi carbon bt i m cc amino acid c cu trc dng D hay L (hnh 3.7) gi l ng phn lp th. S ng phn lp th c tnh theo 2n (n l s carbon bt i) Hu ht cc amino acid khc hp th tia cc tm bc sng () khong t 220 - 280 nm. c bit cng nng 10-3M, trong bc sng khong 280 nm, tryptophan hp th nh sng cc tm mnh nht, gp 4 ln kh nng hp th ca tyrosine (hnh 3.8) v phenylalanine l yu nht.

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Phn ln cc protein u cha tyrosine nn ngi ta s dng tnh cht ny nh lng protein hp ph

- Bc sng(nm)
Hnh 3.8. Ph hp th nh sng cc tm ca tryptophan v tyrosine

3.1.6.4. Tnh lng tnh ca amino acid Trong phn t amino acid c nhm carboxyl -COOH nn c kh nng nhng proton (H+) th hin tnh acid, mt khc c nhm amin- NH2 nn c kh nng nhn proton nn th hin tnh base. V vy amino acid c tnh cht lng tnh. Trong mi trng acid, amino acid dng cation (tch in dng), nu tng dn pH amino acid ln lt nhng proton th nht chuyn qua dng lng cc (trung ho v in), v tip tc tng pH amino acid s nhng proton th hai chuyn thnh dang anion (tch in m). V vy i khi ngi ta coi n nh mt di-acid.

cation

lng cc

anion

Hnh 3.9. Tnh lng tnh ca amino acid

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Tng ng vi phn ly H+ ca cc nhm COOH v NH3+ c cc tr s pK1 v pK2 (biu th phn ly ca cc nhm c 1/2). T trc tin chuyn sang dng lng tnh v sau cng chuyn thnh dng anion.

phn ly ca H+
Hnh 3.10. ng cong chun ca glycine nng 1 M 25OC

Ngi ta xc nh c pHi (pI= pH ng in) = pK1 + pK2 / 2. V d khi ho tan glycine vo mi trng acid mnh th hu nh glycine u dng cation. Nu tng dn lng kim, thu c ng cong chun . Trn ng cong chun thy rng glycine ln lt nhng 2 proton trc Tng ng phn ly ca nhm COOH c mt na c tr s pK1= 2,34 v phn ly ca NH3+ c mt na c tr s pK2= 9,60. Nh vy ta c 2,34 + 9,60 pHi = = 5,97 2

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Bng: 3.2 Cc tr s pK ca cc amino acid thng gp Tn cc amino acid Glycine Alanine Proline Valine Leucine Isoleucine Methionine Phenylalanine Tyrosine Tryptophan Serine Theonine Cysteine Aspargine Glutamine Lysine Histidine Arginine Aspartate Glutamate Cc tr s pK pK1(ca COOH) pK2(ca NH+3) pKR(ca R) 2,34 2,34 1,99 2,32 2,36 2,36 2,28 1,83 2,20 2,38 2,21 2,11 1,96 2,02 2,17 2,18 1,83 2,17 1,88 2,19 9,60 9,60 10,96 9,62 9,60 9,68 9,21 9,13 9,11 9,39 9,15 9,62 10,28 8,80 9,13 8,95 9,17 9,04 9,60 9,67 5,97 6,01 6,48 5,97 5,98 6,02 5,74 5,48 5,66 5,89 5,68 5,87 5,07 5,41 5,65 9,74 7,59 10,76 2,77 3,22 pI

10,07

8,18

10,53 6,00 12,48 3,65 4,25

Mt khc ti pK1 + 2 s phn ly H+ ca nhm COO- glycine l 99%, ch 1% dng COOH v pK2 -2 dng NH3+ l 99%, ch 1% dng NH2. Nh vy trong vng pH t pK1 + 2 n pK2 -2, phn t glycine ch yu dng lng tnh v kt qu ta c mt vng ng in. Ngoi ra cc amino acid trong gc R c thm nhm COOH hay NH2 s phn ly ca chng s c thm mt tr s phn ly na-pKR (xem bng 3.2). 3.1.7. Cc phn ng ho hc ca amino acid Cc amino acid u c nhm NH2 v COOH lin kt vi C, v vy chng c nhng tnh cht ho hc chung. Mt khc cc amino acid khc nhau bi gc R, v vy chng c nhng phn ng ring bit. Ngi ta chia cc phn ng ho hc ca amino acid thnh 3 nhm:

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- Phn ng ca gc R Do cc amino acid c cu to gc R khc nhau, nn ngi ta c th dng xc nh tng amino acid ring r nh phn ng c trng ca n, v d phn ng oxy ho kh do nhm SH ca cysteine, phn ng to mui do cc nhm COOH hay NH2 ca glutamate hay lysine, phn ng to ester do nhm OH ca tyrosine v.v... - Phn ng chung L phn ng c s tham gia ca c hai nhm - COOH v - NH2. Tt c cc amino acid trong phn t protein u phn ng vi hp cht ninhydrin to thnh phc cht mu xanh tm, ring imino acid nh proline to thnh mu vng. Phn ng c thc hin qua mt s bc nh sau: Di tc dng ca ninhydrin nhit cao, amino acid to thnh NH3, CO2 v aldehide, mch polypeptide ngn i mt carbon; ng thi ninhydrin chuyn thnh diceto oxy hindriden. Diceto oxy hindriden, NH3 mi to thnh tip tc phn ng vi mt phn t ninhydrin khc to thnh phc cht mu xanh tm (hnh 3.11)

Hnh 3.11. Phn ng ca protein vi ninhydrin

- Phn ng ring bit C th chia cc phn ng ring bit theo hai nhm - COOH v - NH2 + Cc phn ng ca nhm - COOH. Ngoi cc phn ng ca nhm COOH thng thng to ester, to amid, to mui ...th n cn c nhng phn ng c trng khc nh c th b kh thnh hp cht ru amino di s xc tc ca NaBH4. R-NH2CH-COOH R-NH2CH-CH2OH Nhm COOH c th to thnh phc aminoacyl-adenylate trong phn ng hot ho amino acid tng hp protein, hay c th loi CO2 vn gp rt nhiu trong qu trnh thoi ho amino acid.

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+ Cc phn ng ca nhm - NH2. Nhiu phn ng ca nhm amine c dng nh tnh v nh lng cc ch tiu ca amino acid nh: nh lng nitrogen ca amino acid ngi ta cho phn ng vi HNO2 gii phng N2. R-CH-COOH + HNO2 NH2 R-CH-COOH OH + N2 + H2O

nh lng amino acid ngi ta cho phn ng vi aldehyde to thnh base schiff. xc nh amino acid u N-tn cng ngi ta cho tc dng vi 2-4 dinitrofluobenzen (phn ng Sanger) hay phenyliothiocyanate (phn ng Edman).

3.2. Peptide
3.2.1. Khi nim chung Peptide l nhng protein thng c cu trc on ngn khong t hai n vi chc amino acid ni vi nhau, c khi lng phn t thng di 6.000. Chng c th c tng hp trong t nhin hoc c hnh thnh do s thoi ho protein. Trong cc peptide cc amino acid c lin kt vi nhau thng qua lin kt peptide (hnh 3.12).

Hnh 3.12. S to thnh lin kt peptide

Lin kt peptide c bn cao bi cu trc ca n c 4 e', 2e' thuc v lin kt C=O cn 2e' thuc v b i e' t do ca nguyn t N. Lin kt gia C-N l lin kt phc tp, n c th chuyn t dng n

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dng lai (trung gian) th b mt phn ghp i ca lin kt (hnh 3.13). Ngi ta cho rng t l ca lin kt kp ny l khong 30% i vi lin kt C-N v 70% vi lin kt gia C v O. Nh vy u ca mt chui peptide l amino acid c nhm -amine (-NH2) t do c gi l u Ntn cng v u kia c nhm - carboxyl ( -COOH) t do c gi l u C tn cng. Lin kt peptide to nn b khung chnh ca chui polypeptide, cn cc gc R to nn mch bn ca chui (hnh 3.14)
Dng cng ho tr Dng ion +

Dng lai (hybrid)

Hnh 3.13. S tn ti cc dng ca lin kt peptide

Mch chnh

Mch bn Hnh 3.14. Mch bn v khung ca mt chui polypeptide

3.2.2. Cc phng php xc nh peptide Ngoi phn ng ca nhm NH2 v COOH u tn cng, cc gc R ca peptide cng cho nhng phn ng mu c trng ca cc amino acid t do tng ng. Mt trong nhng phn ng mu c trng nht dnh cho lin kt peptide l phn ng Biure, phn ng ny khng xy ra vi

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amino acid t do v vi dipeptide. Trong mi trng kim mnh, lin kt peptide phn ng vi CuSO4 to thnh phc cht mu tm (hnh 3.15.) v c kh nng hp th cc i bc sng 540 nm. y l phn ng c s dng rng ri nh lng protein. Phng php xc nh protein theo Lowry cng da trn nguyn tc ca phn ng ny bng cch thm thuc th Folin-Ciocalteau lm tng nhy ca phn ng sau khi thc hin phn ng biure, ng thi da vo cc gc Tyr, Try nh thuc th to phc mu xanh da tri. OC =NH HN C NH HN O Cu O C OHN = C NH

Hnh 3.15. Phc mu tm trong phn ng Biure

C mt s phng php tch phn lp v xc nh thnh phn, s lng v trnh t amino acid trong peptide. V nguyn tc chung cc phng php tch phn lp v xc nh peptide v c bn cng nh i vi protein. Tuy nhin peptide l nhng on ngn ca chui polypeptide, v th c th b qua giai on ct chui polypeptide thnh cc peptide nh m c th tch, phn lp ngay bng phng php in di hay sc k tch ring tng peptide. Sau khi tch ring cc peptide, tin hnh thu phn hon ton thnh cc amio acid t do, xc nh cc amino acid , amino acid u N-tn cng v amino acid u C-tn cng. Cc d liu thu c qua s phn tch ny s c so snh i chiu v tng hp li. V d, Puppy v Bodo phn tch mt peptide ca dch khi thu phn Cytocrom C thu c cc d kin sau y: - Thnh phn amino acid ca peptide sau khi c thu phn hon ton v tin hnh sc k l 2Cys, 1 Ala, 2 Glu, 1His, 1Thr, 1Val,v 1Lys.

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- Dng phng php Sanger xc nh c amino acid u N-tn cng l Cys v phng php carboxypeptidase xc nh c amino acid u C - tn cng l Lys. - Cu to ca peptide nh (bng cch thu phn tng phn ban u v xc nh cc amino acid, amino acid u N - tn cng v amino acid u C - tn cng ca mi peptide nh): Cys- Ala Cys-(Ala,Glu) Ala- Glu Glu- Cys Cys- His Glu (Cys, His) (Val- Glu) Thr (Val, Glu) Glu- Lys Thr (Val, Glu, Lys) Tng hp cc d kin trn, h xc nh c trnh t cc amino acid ca peptide nghin cu l: H2N-Cys-Ala-Glu-Cys-His-Thr-Val-Glu-Lys-COOH. y l nguyn tc chung xc nh mt trnh t trong peptide. Tuy nhin i vi nhng peptide di, vic xc nh rt phc tp. 3.2.3. Cc peptide thng gp trong thin nhin Trong t nhin tn ti nhiu dng peptide c chc phn quan trong lin quan n hot ng sng ca c th nh l cc hormon, cc cht khng sinh hay nhng cht tin thn ca t bo vi khun v.v... Bn cnh cng c nhng peptide chc phn cha r rng, c nhng peptide l sn phm thu phn ang cn dang d ca protein. Trong phm vi ca gio trnh ny xin c gii thiu mt s peptide quan trng,c nhiu ngha i vi cho hot ng sng ca sinh vt. 3.2.3.1. Glutathion v cc cht tng t Glutathion l mt tripeptide -glutamyl-cysteyl-glycine c cng thc cu to nh sau: NH2 CH2 SH

HOOC-CH-CH2-CH2-CO-NH-CH-CO-NH-CH2-COOH Trong cu trc ca glutathion, nhm SH ca cysteine l nhm hot ng, v vy ngi ta thng vit tt ch glutathion l G-SH. y l glutathion dng kh. Trong mi trng hot ng glutathion c th

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nhng hydrogen (H) thnh dng oxy ho (GSSG) v ngc li c th nhn H thnh dng kh: 2GSH
-2H +2H

G-S-S-G

Nh phn ng trn, glutathion ng vai tr ca mt h thng oxy ho kh (vn chuyn hydrogen). Glutathion l mt trong nhng peptide ni bo ph bin nht, n phn b nhiu trong cc m v cc c quan nh: gan, thn, lch, tim, phi, hng cu v.v... 3.2.3.2. Cc hormone sinh trng (HGH) Hormone sinh trng ca ngi (HGH-human growth hormone) cn c tn gi STH (somatotropin hormone) l mt chui polypeptide bao gm 191 amino acid c khi lng phn t 20.000. Trong cu trc c hai cu disulfua c to thnh gia amino acid 53 -165 v gia amino acid 182-189. Hot ng sinh hc ca HGH l chui gm 134 amino acid. HGH c cu to rt ging vi hormon lactogen ca rau thai (85% amino acid ging nhau) v gn ging prolactin ca ngi (32% amino acid ging nhau). Hormon sinh trng c tc dng i vi s tng trng ni chung, kch thch s to sn hn l to xng, n cng l mt hormon chuyn ho. Hormon sinh trng kch thch s tng hp protein t nhng amino acid c vn chuyn d dng vo trong t bo nh chng, v l hormon gy tng ng huyt, sinh i tho ng, ng thi kch thch s thoi ho lipid m bo nhu cu v nng lng cho c th, gy tng acid bo t do trong huyt tng. S thiu ht HGH nu xy ra trc tui dy th s dn n chng ngi ln, s d tha HGH nu xy ra trc tui dy th s dn n chng ngi khng l, nu xy ra sau tui dy th s dn n chng ngi b to cc (pht trin chiu dy ca u, xng v mt).

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3.2.3.3. Insulin T 1953, Sanger (gii thng Nobel 1958) nghin cu, tinh ch v xc nh hon ton cu trc ca phn t insulin. Phn t insulin bao gm 51 amino acid, c cu trc gm 2 chui polypeptide, vi khi lng phn t 5.700 chui A c 21 amino acid, chui B c 30 amino acid. Hai chui c ni vi nhau bng 2 cu disulfua. Trong chui A cng hnh thnh 1 cu disulfua gia amino acid th 6 v amino acid th 11. Phn c hiu (c trng ca mt loi) ch tp trung vo cc amino acid th 8-9-10, 12-14 ca chui A v c bit l amino acid th 30 ca chui B (hnh 3.16). Ngi ta cng xc nh c cu trc ba chiu ca insulin v thy rng cu trc phn t insulin c gi vng bi nhiu lin kt mui, lin kt hydro v lin kt cu disulphate gia chui A v chui B. Insulin c tc dng r nht trong tt c cc hormon ca tuyn tu, c bit i vi qu trnh chuyn ho glucid, n c tc dng h ng huyt. Insulin cn kch thch qu trnh tng hp v c ch qu trnh thoi ho glycogen c, gan v m m. c bit, insulin tng cng tng hp acid bo, protein v kch thch s ng phn. Tc dng quan trng nht ca insulin l kch thch s thm nhp glucose, mt s ose, amino acid trogn t bo c v m. Do vy insulin lm gim lng glucose trong mu. Ngoi ra insulin cng lm gim s tn to glucose do lm gim nng enzyme nh pyruvat carboxylase v fructose 1-6 diphosphatase.

Hnh 3.16. Cc amino acid ca chui A v B insulin b

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3.2.3.4. Oxytocin, Vasopressin Vasotocin Oxytocin l mt peptide c 9 amino acid. ng vt c v, oxytocin ch khc s thay i ca 2 amino acid l: amino acid v tr th ba l isoleucine v amino acid v tr th tm l leucine (bng 3.3). Vasopressin ca loi ch nhi c cu trc trung gian gia vasopresin v oxytocin ca ng vt c v (amino acid th ba l isoleucin v amino acid th tm l arginine v c tn l vasotocin). Vasopressin l mt peptide c cu trc gm 9 amino acid. .
Bng 3.3. So snh cu trc ho hc gia oxytocin v vasopressin ca mt s loi ng vt VaLysine Vasopressin 1 2 3 4 5 6 7 8 9 Cys-Tyr-Phe-Glu-Asn-Cys-Pro-Lys-Gly-NH2 Ln, H m Phn ln ng vt c v

So2 3 4 5 6 7 8 9 Arginine 1 vasopressin Cys-Tyr-Phe-Glu-Asn-Cys-Pro-Arg-Gly-NH2 pres1 2 3 4 5 6 7 8 9 Cys-Tyr-Ile-Glu-Asn-Cys-Pro-Arg-Gly-NH2 1 2 3 4 5 6 7 8 9 Cys-Tyr-Ile-Glu-Asn-Cys-Pro-Leu-Gly-NH2

sin

Vasotocin

ng vt c xng sng, khng c v ng vt c xng sng c v, chim

Oxytocin

Phn ln ng vt c v amino acid th 8 ca vasopressin l arginine (arg-vasopressin), tr ln v h m, amino acid th 8 l lysine (lysvasopressin). Oxytocin c tc dng trn c trn ca t cung v tuyn v, gy co khi t cung sinh con v kch thch s tit sa khi cho con b. Vasopressin c tc dng chng li niu, tng cng ti hp thu nc thn, ng thi lm co mch, do c tc dng tng huyt p.

3.3. Protein
3.3.1. Cu trc phn t bc 1, 2, 3, 4 V mt cu trc ngi ta phn bit protein gm bn bc: bc I, bc II, bc III v bc IV (Hinh. 3.17)

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Bc I Bc II Bc III Bc IV

Hnh 3.17. S cc bc cu trc ca protein

3.3.1.1. Cu trc bc I Cu trc bc I biu th trnh t cc gc amino acid trong chui polypeptide, cu trc ny c gi vng bng lin kt peptide (lin kt cng ha tr). Cu trc bc I l phin bn ca m di truyn, vic xc nh c cu trc bc I l c s tng hp nhn to protein bng phng php ho hc hoc bng k thut ca cng ngh sinh hc. Hin nay nhiu loi protein bit c trnh t cc amino acid trong chui polypeptide nh: ribonuclease l mt protein c 124 amino acid c ni vi nhau thnh mt chui, c 4 cu disulfua (hnh 3.18); hemoglobin l protein c 4 chui polypeptide, 2 chui ( mi chui 141 amino acid) v 2 chui (mi chui 146 amino acid); trypsinogen b (229 amino acid); kimotrypsin b (229 amino acid); alcol dehydrogenase nga (374 amino acid); glutamate dehydrogenase b (500 amino acid) v.v..

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Hnh 3.18. Cu trc bc nht ca ribonuclesae ca b

3.3.1.2. Cu trc bc II Biu th s xon ca chui polypeptide, l tng tc khng gian gia cc gc amino acid gn nhau trong mch polypeptide.

Xon

Lin kt hydrogen Xon Hnh 3.19. Cc kiu xun trong cu trc bc II ca protein

Ni cch khc, cu trc bc II l dng khng gian cc b ca tng phn trong mch polypeptide. Cu trc ny c lm bn nh cc lin kt

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hydrogen c to thnh gia lin kt peptide k gn nhau, cch nhau nhng khong xc nh. Theo Pauling v Cori (1951) cu trc bc II ca protein bao gm 2 kiu chnh l xon v phin gp . trong tc ngi ta tm thy keratin l loi protein c hai dng cu trc: dng bnh thng v dng dui thng.; cu trc phin gp tm thy trong fibroin ca t. Cu trc xun hin nay c tm thy trong nhiu loi protein khc nhau Mt khc t l % xon trong cc protein khc nhau cng thay i kh nhiu. V d trong hemoglobin v mioglobin l 75%; lysosyme l 35%; ribonuclease l 17% ..
Bng 3.4. S lng xon v phin gp trong chui n mt s protein S gc (%) Protein (s gc) Chymotrypsin (247) Ribonuclease (124) Carboxypeptidase (397) Cytochrom C (104) Lysosyme (129) Myoglobin (153) Xon 14 26 38 39 40 78 Phin gp 45 35 17 0 12 0

Ngoi ra cn c kiu xon collagen c tm thy trong phn t collagen (hnh 3.20).

Hnh 3.20. Cu trc kiu xun collagen

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n v cu trc ca n l tropocollagen bao gm 3 mch polypeptide bn vo nhau thnh mt dy cp siu xon (v mi mch n c cu trc xon, chiu cao ca mi gc xon trn trc siu xon ny l 2,9 anstron, mt vng xon l 3,3 gc amino acid. Ba chui polypeptide trong dy cp ni vi nhau bng cc lin kt hydrogen. 3.3.1.3. Cu trc bc III Biu th s xon v cun khc ca chui polypeptide thnh khi, c trng cho potein cu, l tng tc khng gian gia cc gc amino acid xa nhau trong chui polypeptide. Trong nhiu protein hnh cu c cha cc gc Cys to nn lin kt disulfua gia cc gc Cys xa nhau trong chui polypeptide lm cho chui b cun li (xem myoglobin hnh 3.21). Ngoi ra cu trc bc III cn c gi vng bng cc loi lin kt khc nh Van der Waals, lin kt hydrogen, lin kt tnh in gia cc gc amino acid v.v... 3.3.1.4. Cu trc bc IV Biu th s kt hp ca cc chui c cu trc bc III trong phn t protein. Hay ni cch khc, nhng phn t protein c cu trc t 2 hay nhiu chui protein hnh cu, tng tc vi nhau trong khng gian to nn cu trc bc IV. Mi mt chui polypeptide c gi l mt tiu n v (subunit), chng gn vi nhau nh cc lin kt hydrogen, tng tc Van der Waals gia cc nhm phn b trn b mt ca cc tiu n v lm bn cu trc bc IV.

Myoglobin

Hemoglobin

Hnh 3.21. Cu trc bc III ca myoglobin v bc IV ca hemoglobin

(hemoglobin l protein c 4 chui polypeptide2 chui v 2 chui ; myoglobin ch gm mt chui polypeptide)

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3.3.2. Mt vi tnh cht ca protein 3.3.2.1. Tnh cht l-ho ca protein - Tnh tan ca protein Cc loi protein khc nhau c kh nng ho tan d dng trong mt s loi dung mi nht nh, chng hn nh albumin d tan trong nc; globulin d tan trong mui long; prolamin tan trong ethanol, glutelin ch tan trong dung dch kim hoc acid long v.v... - Tnh ngm nc ca protein Trong mi trng nc, protein kt hp vi nc trng ln tr thnh dng keo, hay ni cch khc protein trng thi hydrate ho, cc phn t nc bm vo cc nhm a nc trong phn t protein nh -NH2, -COOH..., lp o nc bao quanh phn t protein l mt trong cc yu t lm bn vng cu trc, ngn cch cc phn t protein khng cho chng dnh vo nhau thnh ta. - nht ca dung dch protein Khi protein ho tan trong dung dch, mi loi dung dch ca nhng protein khc nhau c nht khc nhau (bng 3.5). Ngi ta c th li dng tnh cht ny xc nh khi lng phn t ca protein ( nht cng cao th khi lng phn t cng cao).
Bng 3.5. nht ca mt s protein Protein Gelatin Albumin trng Gelatin Albumin trng Nng % (trong nc) 3,0 3,0 8,0 8,0 nht tng i (ca nc =1) 4,54 1,20 14,2 1,57

- Hng s in mi ca dung dch protein Khi thm cc dung mi hu c trung tnh nh ethanol, aceton vo dung dch protein trong nc th tan ca protein gim v protein s kt ta do vic gim mc hydrate ho ca cc nhm ion ho ca protein, lp o mt nc, cc phn t protein kt hp vi nhau thnh ta. Nh vy, hng s in mi ca dung mi lm ngn cn lc tnh in gia cc nhm tch in ca protein v nc. Mi lin h c c trng bi biu thc:

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L1 - l2 F= D - hng s in mi ca dung dch F- lc tnh in gia cc ion tch in L1 , l2 - in tch cc ion, r - khong cch gia cc ion - Tnh cht in li ca protein Cng nh cc amino acid, protein l cht in li lng tnh v trong phn t protein c nhiu nhm phn cc mnh (bn gc R) ca amino acid v d: nhm COOH th hai ca Asp, Glu; nhm NH2 ca Lys; nhm OH ca Ser, Thr, Tyr v.v...Trng thi tch in ca cc nhm ny ph thuc vo pH ca mi trng. mt pH no m tng in tch (+) v in tch (-) ca phn t protein bng khng, phn t protein khng di chuyn trong in trng gi l pHi (isoelectric - im ng in) ca protein. Nh vy protein cha nhiu Asp, Glu (amino acid c tnh acid mnh) th pHi trong vng acid, ngc li nhiu amino acid kim nh Lys, Arg, His th pHi trong vng kim. mi trng c pH < pHi , protein a s l mt cation, s in tch dng ln hn s in tch m. pH > pHi phn t protein th hin tnh acid, cho ion H+, do s in tch m ln hn s in tch dng, protein l mt a anion, tch in m.
Bng 3.5. Gi tr pHi ca mt s protein Protein Pepsin Albumin trng Casein Albumin huyt thanh Gelatin pHi 1,0 4,6 4,7 4,9 4,9 Protein Globulin sa Hemoglobin Ribonuclease Trypsin Cytochrom C Prolamin pHi 5,2 6,8 7,8 10,5 10,6 12,0

Dr2 Trong :

Trong mi trng c pH = pHi ca protein, protein d dng kt t li vi nhau v th ngi ta li dng tnh cht ny xc nh pHi ca protein cng nh kt ta protein. Mt khc do s sai khc nhau v pHi gia cc protein m c th iu chnh pH ca mi trng tch ring cc protein ra khi hn hp ca chng.

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- S kt ta bng mui ca dung dch protein Mui trung tnh c nh hng r ti ho tan ca protein hnh cu: vi nng thp chng lm ho tan nhiu protein. Tc dng khng ph thuc vo bn cht ca mui trung tnh, m ph thuc vo nng mui v s in tch ca mi ion trong dung dch, tc l ph thuc vo lc ion ca dung dch ( = 1/2 C1 Z1, 2 trong l k hiu ca tng, C1 l nng ca mi ion, Z1 l in tch ca mi ion). Cc mui c ion ho tr 2 (MgCl2, MgSO4...) lm tng ng k tan ca protein hn cc mui c ion ho tr 1 (NaCl, NH4Cl, KCl...). Khi tng ng k nng mui trung tnh th tan ca protein bt u gim v nng mui rt cao, protein c th b kt ta hon ton. Cc protein khc nhau b kt ta nhng nng mui trung tnh khc nhau. Ngi ta s dng tnh cht ny chit xut v tch ring protein khi hn hp. l phng php dim tch (kt ta protein bng mui). Th d dng mui amonium sulfate 50% bo ho kt ta globulin v dung dch amonium sulfate bo ho kt ta albumin t huyt thanh. - Biu hin quang hc ca protein Cng nh nhiu cht ho hc khc, protein c kh nng hp th v bc x nh sng di dng lng t h. V vy c th o cng hp th ca protein trong dung dch hay cn gi l mt quang thng k hiu bng ch OD (Optical Density). Da trn tnh cht ngi ta sn xut ra cc loi my quang ph hp th phn tch protein. Nhn chung protein u c kh nng hp th nh sng trong vng kh kin (t 350nm- 800nm) v vng t ngoi (t 320nm xung ti 180nm). Trong vng nh sng kh kin protein kt hp vi thuc th hp th mnh nht vng nh sng 750nm (nh lng protein theo Lowry). i vi vng t ngoi dung dch protein c kh nng hp th nh sng t ngoi hai vng bc sng khc nhau: 180nm-220nm v 250nm - 300nm. bc sng t 180nm-220nm l vng hp th ca lin kt peptide trong protein, cc i hp th 190nm. Do lin kt peptide c nhiu trong phn t protein nn hp th kh cao, cho php nh lng tt c cc loi protein vi nng thp. Tuy nhin vng hp th ny ca cc lin kt peptide trong protein c th b dch v pha c bc sng di hn khi c mt s tp cht ln trong dung dch protein. Mt khc chnh cc tp cht ny cng hp th nh sng t ngoi vng bc sng 180nm-220nm. V th trong thc t thng o hp th ca dung dch protein bc sng 220nm-240nm. bc sng t 250nm-300nm l vng hp th cc amino acid thm (Phe, Tyr, Trp) c trongphn t protein hp th cc i 280nm. C th s dng phng php o hp th ca dung dch protein bc sng 280nm

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nh tnh v nh lng cc protein c cha cc amino acid thm. Hm lng cc amino acid thm trong cc protein khc nhau thay i kh nhiu, do dung dch ca cc protein khc nhau c nng ging nhau c th khc nhau v hp th bc sng 280nm. Ngoi ra c nhiu cht khc trong dung dch cng c nh hng n hp th protein. V vy, cc phng php o hp th vng nh sng t ngoi thng c dung nh lng protein c tinh sch hoc xc nh protein trong cc phn on nhn c khi sc k tch cc protein qua ct. - Kt ta thun nghch v khng thun nghch protein Khi protein b kt ta n thun bng dung dch mui trung tnh c nng khc nhau hoc bng alcohol, acetone nhit thp th protein vn gi nguyn c mi tnh cht ca n k c tnh cht sinh hc v c th ho tan tr li gi l kt ta thun nghch. Cc yu t kt ta thun nghch c dng thu nhn ch phm protein. Trong qu trnh kt ta thun nghch mui trung tnh va lm trung ho in va loi b lp v hydrate ho ca protein, cn dung mi hu c vn ho nc s ph hy lp v hydrate nhanh chng. Trong ch phm protein nhn c cn ln cc cht dng kt ta, cn s dng phng php thch hp loi b cc cht ny. V d c th dng phng php thm tch loi b mui. Ngc li kt ta khng thun nghch l protein sau khi b kt ta khng th phc hi li trng thi ban u. S kt ta ny thng c s dng loi b protein ra khi dung dch, lm ngng phn ng ca enzyme. Mt trong nhng yu t gy kt ta khng thun nghch n gin nht l un si dung dch protein (s ni k hn trong phn bin tnh protein sau). - Cc phn ng ho hc ca protein Cng nh cc amino acid v peptide, protein c cc phn ng ho hc tng t, l: phn ng ca cc nhm -COOH, -NH2, gc R v phn ng to mu c trng ca lin kt peptide nh phn ng biure (xem chng 2 v 3. y xin c gii thiu thm mt s phn ng mu c trng khc, c ngha quan trng trong pht hin protein v cc gc amio acid trong chui polypeptide. + Phn ng vi thuc th Folin-Ciocateau Thuc th Folin-Ciocateau c cha phosphomolipdic acid v phosphovolframic acid cc cht ny lm tng nhy ca phn ng biure, mt khc phn ng vi gc Tyr v Trp trong phn t protein. Cc gc amino acid ny tham gia trong qu trnh to phc cht mu xanh da tri. + Cc phn ng mu c trng khc ca protein

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Nhng phn ng ny c c l do s c mt ca cc nhm nh chc ha hc xc nh trong phn t protein. C th s dng chng pht hin amino acid, protein trong dung dch. Phn ng xanthproteic: cc gc amino acid Tyr, Trp, Phe trong protein tc dng vi HNO3 c to thnh mu vng v sau khi thm kim s chuyn thnh da cam. Phn ng Pauli: cc gc Tyr, His trong protein tc dng vi diazobenzosulfonic acid to thnh mu anh o. Phn ng Millon: gc Tyr tc dng vi thu ngn nitrate trong HNO3 c to thnh kt ta mu nu t. Phn ng Saccaguichi: gc Arg tc dng vi dung dch kim ca -naphtolv hypobromitecho mu anh o. Phn ng Adamkievich: gc Trp tc dng vi glyoxylic acid v H2SO4 c to thnh vng tm mt phn cch. 3.3.2.2. Tnh cht v chc nng sinh hc ca protein Protein l hp cht hu c c ngha quan trng bc nht trong c th sng, v mt s lng, n chim khng di 50% trng lng kh ca t bo. T lu, bit rng protein tham gia mi hot ng sng trong c th sinh vt, ngoi vai tr l thnh phn chnh trong cu trc ca t bo v m, protein cn c nhiu chc nng phong ph khc quyt nh nhng c im c bn ca s sng nh s truyn t thng tin di truyn, s chuyn ho cc cht do cc enzyme, cc khng th chng li bnh tt, cc hormon dn truyn cc tn hiu trong t bo v.v... u c bn cht l cc protein. Trong c th protein c mt s vai tr sau y: - Chc nng to hnh Ngoi cc protein lm nhim v cu trc nh v virus, mng t bo, cn gp nhng protein thng c dng si nh: sclerotin c trong lp v ngoi ca su b; fibroin ca t tm, nhn; collagen, elastin ca m lin kt, m xng. Collagen m bo cho bn v tnh mm do ca m lin kt. - Chc nng xc tc Hu ht cc phn ng xy ra trong c th u do cc protein c bit ng vai tr xc tc, nhng protein c gi l cc enzyme. Mc d gn y ngi ta pht hin c mt loi RNA c kh nng xc tc qu trnh chuyn ho tin RNA thng tin (pre-mRNA) thnh RNA thng tin (mRNA), ngha l enzyme khng nht thit phi l protein. Nhng nh ngha c tnh cht kinh in: enzyme l nhng protein c kh nng xc tc

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c hiu cho cc phn ng ho hc, l cht xc tc sinh hc vn c ngha c bit quan trng. Hin nay ngi ta bit c khong 3.500 enzyme khc nhau, nhiu enzyme c tinh sch, kt tinh v nghin cu cu trc. - Chc nng bo v Ngoi vai tr l thnh phn chnh trong xc tc, cu trc ca t bo v m, protein cn c chc nng chng li bnh tt bo v c th. l cc protein tham gia vo h thng min dch, c bit nhiu loi protein thc hin cc chc nng ring bit to nn hiu qu min dch c hiu v khng c hiu. Cc protein min dch c nhc n nhiu hn c l cc khng th, b th v cc cytokine. Ngoi ra protein cn tham gia vo qu trnh ng mu chng mt mu cho c th hoc mt s loi c th sn xut ra nhng c t c bn cht l protein nh enzyme nc rn, lectin v.v...,c kh nng tiu dit k th bo v c th. - Chc nng vn chuyn Trong c th c nhng protein lm nhim v vn chuyn nh hemoglobin, mioglobin, hemocyanin vn chuyn O2, CO2 v H+ i khp cc m, cc c quan trong c th. Ngoi ra cn c nhiu protein khc nh lipoprotein vn chuyn lipid, ceruloplasmin vn chuyn ng (Cu) trong mu v.v...Mt trong nhng protein lm nhim v vn chuyn c nhc n nhiu nht l hemoglobin. - Chc nng vn ng. Nhiu protein lm nhim v vn ng co rt nh myosin, actin si c, chuyn v tr ca nhim sc th trong qu trnh phn bo v.v... -Chc nng d tr v dinh dng Cc protein lm nhim v d tr nh casein ca sa, ovalbumin ca trng, feritin ca lch (d tr st) v.v...Protein d tr ny chnh l ngun cung cp dinh dng quan trng cho cc t chc m, phi pht trin. - Chc nng dn truyn tn hiu thn kinh Nhiu loi protein tham gia vo vic dn truyn tn hiu thn kinh i vi cc kch thch c hiu nh: sc t th gic rodopsin mng li mt. - Chc nng iu ho Cc protein lm nhim v iu ho qu trnh trao i cht thng qua tc ng ln b my thng tin di truyn nh cc hormon, cc protein c ch c hiu enzyme u c chc nng iu ho nhiu qu trnh trao i cht, chng hn cc protein repressor vi khun c th lm ngng qu trnh sinh tng hp enzyme t cc gen tng ng.

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- Chc nng cung cp nng lng Protein cng l ngun cung cp nng lng quan trng cho mi hot ng sng trong c th. Trong c th cc protein b phn gii thnh cc amino acid, t tip tc to thnh hng lot cc sn phm, trong c cc ceto acid, aldehyde v carboxylic acid. Cc cht ny u b oxy ho dn dn to thnh CO2 v H2O ng thi gii phng ra nng lng. 3.3.3. Phn loi protein Protein l hp cht a dng v thnh phn cu trc, chc nng; khi lng phn t ca chng cng rt khc nhau (bng 3.6). Do s phn loi protein c th da vo nhng mc ch khc nhau. C hai cch phn loi thng thng nht: 3.3.3.1. Phn loi theo hnh dng - Protein dng si C hnh dng di, thng l hnh si, chiu di ca phn t protein si ln hn ng knh ca n hng trm ln. Protein si tng i bn vng, khng tan trong nc v dung dch mui long, cc chui polypeptide ca protein si nm dc theo mt trc thnh nhng si di. Protein si l yu t cu trc c bn ca m lin kt ng vt cao cp, th d collagen gn v m xng,elastin m lin kt n hi, -keratin tc da v.v... - Protein dng cu C dng gn nh hnh cu hoc hinh bu dc, chiu di ca phn t protein cu c th ln hn ung knh ca n t 3 n 10 ln. Protein cu khng bn vng bng protein si, a s tan trong dung dch nc v d khuych tn, thng c chc nng hot ng sng ca t bo nh cc enzyme, cc hormon, cc protein vn chuyn nh albumin huyt thanh, hemoglobin v.v... - Protein dng trung gian Mt s protein dng trung gian, va c c im ca protein si, va c c im ca protein cu. Th d myosin (yu t cu trc v chc nng quan trong ca c) c cu trc hnh que di l c im ca protein si, nhng li tan trong dung dch mui l c im ca protein cu. Ngoi ra, trong nhm trung gian ny cn c cht tin thn ca fibrin l fibrinogen. 3.3.3.2. Phn loi theo thnh phn ho hc Protein gm hng trm, hng ngn, thm ch hng vn amino acid ni vi nhau bng lin kt peptide to nn mt hay nhiu chui polypeptide c cu trc rt phc tp.

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Cn c s c hay vng mt ca mt s thnh phn c bn cht khng phi protein m ngi ta chia protein thnh hai nhm:
Bng: 3.6 Khi lng (MW) v cu trc phn t ca mt s protein Protein Glucagon Insulin Ribonuclease (ty b) Lysozyme (lng trng trng) Myoglobin (tim nga) Chymotripsin (ty b) Hemoglobin (ngi) Albumin (huyt thanh ngi) Hexokinase (men bia) Tryptophan-synthetase (E.coli) -globulin (nga) Glycogen-phosphorylase (c th) Glutamate-dehydrogenase (gan b) Synthetase ca acid bo (men bia) Virus khm thuc l Khi lng (Dalton) 3482 5733 12.640 13.930 16.890 22.600 64.500 68.500 96.000 117.000 149.000 495.000 1.000.000 2.300.000 40.000.000 S gc amino acid 29 51 124 129 153 241 574 550 800 975 1.250 4.100 8.300 20.000 336.500 S chui polypeptide 1 2 1 1 1 3 4 1 4 4 4 4 40 21 2.130

- Protein n gin Protein n gin l nhng protein m phn t ca chng gm ton amino acid. Th d mt s enzyme ca tu b nh ribonuclease gm ton amino acid ni vi nhau thnh mt chui polypeptide duy nht (c 124 gc amino acid, khi lng phn t 12.640), chymotrypsin gm ton amino acid ni vi nhau thnh ba chui polypeptide (c 241 gc amino acid, khi lng phn t 22.600) v.v...Da theo kh nng ho tan trong nc hoc trong dung dch ngi ta c th chia cc protein n gin ra mt s nhm nh nh: - Albumin: tan trong nc, b kt ta nng mui (NH4)2SO4 kh cao (70-100%). - Globulin: khng tan hoc tan t trong nc, tan trong dung dch long ca mt s mui trung tnh nh NaCl, KCl, Na2SO4..., v b kt ta nng mui (NH4)2SO4 bn bo ho. - Prolamin: khng tan trong nc hoc dung dch mui long, tan trong ethanol, isopropanol 70-80%.

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- Glutein: ch tan trong dung dch kim hoc acid long. - Histon: l protein c tnh kim d tan trong nc, khng tan trong dung dch amoniac long. - Protein phc tp Protein phc tp l nhng protein m phn t ca chng ngoi cc amino acid nh protein n gin cn c thm thnh phn khc c bn cht khng phi l protein cn gi l nhm thm (nhm ngoi). Tu thuc vo bn cht ca nhm ngoi m ngi ta chia cc protein phc tp ra cc nhm nh v thng gi tn cc protein bt u bng tip u ng ch bn cht nhm ngoi: - Lipoprotein: nhm ngoi l lipid. - Nucleoprotein: nhm ngoi l nucleic acid. - Glucoprotein: nhm ngoi l carbohydrate v dn xut ca n. - Phosphoprotein: nhm ngoi l phosphoric acid. - Chromoprotein: nhm ngoi l hp cht c mu. Tu theo tnh cht ca tng nhm ngoi m c nhng mu sc khc nhau nh ( hemoglobin), vng ( flavoprotein)...

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TI LIU THAM KHO

Ti liu ting Vit
1. Trn Th n, i Duy Ban, Nguyn Hu Chn, nh H, L c Trnh. 1980. Ho sinh hc. NXB Y hc, H Ni 2.Phm Th Trn Chu, Trn Th ng. 1999. Ho sinh hc. NXB Gio dc, H Ni. 3. Nguyn Ln Dng, Phm Vn Ty, Nguyn nh Quyn. 1999. Vi sinh vt hc. NXB Gio dc,H Ni. 4. L c Trnh. 1998. Hormon. NXB Y hc. H ni

Ti liu ting Anh

1. Dennison C. 2002. A Guide To Protein Isolation. Kluwer Academic Publishers. New York, Boston, Dordrecht, Lodon, Moscow. 2. Fersht A., 1998, Structure and Mechanism in Protein Science, W. H. Freeman, 3rd Rev Edit. 3. Lehninger A. L. 2004. Principle of Biochemistry, 4th Edition. W.H Freeman, 2004 4. Liebler D. C. 2002. Introduction to proteomics. Humana Press Inc. Totuwa, New Jersey. 5. Lodish H, 2003. Molecular Cell Biology. 5th ed, W.H Freeman, 2003 6. Walker J. M. 1996. The Protein Protocols Hand book. 2nd ed. Humana Press Inc. Totuwa, New Jersey.