Bi ging

CNG NGHE PROTEIN

GS 1S Cao ng Nguyn
Chuong 1 M U
I. Khi qut chung v protein
11 Nb J"c tt cbo c%o bm cb!t ptotel
- V mt s luong, n chim khng dui 50% trong
luong kh ca t bo.
-V thnh phn cu trc, protein duoc tao thnh ch
yu t cc amino acid qua lin kt peptide.
- Cho dn nay ngui ta d thu duoc nhiu loai
protein dang sach cao c th kt tinh duoc v d
xc djnh duoc thnh phn cc nguyn t ho hoc,
thng thung trong cu trc ca chng 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 djnh.
- Nh tnh cht ny d djnh luong protein theo
phuong php Kjeldahl, ngui ta tnh luong N ri
nhn vi h s 6,25.
- Ngoi ra trong protein cn gp mt s nguyn t
khc nhu S 0-3% v P, Fe, Zn, Cu...
Khi luong phn t, k hiu l Mr (duoc tnh bng
Dalton)* ca cc loai protein thay di trong nhng gii
han rt rng, thng thung t hng trm cho dn hng
triu.
V du: insulin c khi luong phn t bng 5,733.
glutamat-ehydrogengenase trong gan b c khi luong
phn t bng 1.000.000 (bng 1.1).
1.2. ngha khoa hc v thc tin c%a nhm ch!t
protein
T lu, d bit rng protein tham gia moi hoat dng
sng trong co th sinh vt, t vic tham gia xy dung t
bo, m, dn tham gia hoat dng xc tc v nhiu chc
nng khc v.v...
II. Phn Ioai protein
- Protein gm hng trm, hng ngn amino acid
ni vi nhau bng lin kt peptide tao nn mt hay
nhiu chui polypeptide c cu trc rt phc tap.
- Cn c su c mt hay vng mt ca mt s
thnh phn c bn cht khng phi protein m ngui ta
chia protein thnh hai nhm ln:
2.1. Protein don gin
Protein don gin l nhng phn t m thnh phn cu
tao ca n gm hon ton amino acid. Th du mt s
enzyme ca tuy b nhu ribonuclease gm hon ton
amino acid ni vi nhau thnh mt chui polypeptide
duy nht (c 124 gc amino acid, khi luong phn t
12.640), chymotripsin gm ton amino acid ni vi
nhau thnh chui polypeptide (c 241 gc amino acid,
khi luong phn t 22.600)v.v...
Dua theo kh nng ho tan trong nuc hoc trong dung djch
dm mui, kim hoc dung mi hu co ngui ta c th chia cc
protein don gin ra mt s nhm nh nhu:
-Albumin: tan trong nuc, bj kt ta nng d mui (NH
4
)
2
SO
4
kh cao (70-100%).
-Globulin: khng tan hoc tan t trong nuc, tan trong dung djch
mui long ca mt s mui trung tnh nhu NaCl, KCl, a
2
SO
4
...,
v bj kt ta nng d mui (NH
4
)
2
SO
4
bn bo ho.
-Prolamin: khng tan trong nuc hoc dung djch mui long, tan
trong ethanol, isopanol 70-80%.
-Glutein: chi tan trong dung djch kim hoc acid long.
-Histon: l protein c tnh kim d tan trong nuc, khng tan
trong dung djch amoniac long.
2.2. Protein phc tp
Protein phc tap l nhng protein m thnh phn
phn t ca n ngoi cc -- amino acid nhu protein don
gin cn c thm thnh phn khc c bn cht khng phi
l protein cn goi l nhm thm (nhm ngoai).
Tuy thuc vo bn cht ca nhm ngoai, ngui ta
chia cc protein phc tap ra cc nhm nh v thung goi
tn cc protein d theo bn cht nhm ngoai:
-Lipoprotein: nhm ngoai l lipide.
-Nucleoprotein: nhm ngoai l acid nucleic.
-Glycoprotein: nhm ngoai l carbohydrate v dn xut ca
n.
-Phosphoprotein: nhm ngoai l acid phosphoric.
-Cromoprotein: nhm ngoai l hop cht c mu. Tuy theo
tnh cht ca tng nhm ngoai m c nhng mu sc khc
nhau nhu d ( hemoglobin), vng ( flavoprotein)...
III. Chc nng sinh hc ca protein
.1. Xc tc v enzyme
3.1.1. Quan dim v xc tc enzyme
Hu ht tt c cc phn ng xy ra trong co th du do cc
protein dc bit dng vai tr xc tc, nhng protein d duoc
goi l cc enzyme. Mc d gn dy ngui ta d pht hin
duoc mt loai 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 enzyme xc tc sinh hoc c bn cht l acid
nucleic duoc goi l ribozyme. Nhung djnh ngha c tnh cht
kinh din: enzyme l nhng protein c kh nng xc tc dc
hiu cho cc phn ng ho hoc, l cht xc tc sinh hoc vn
c ngha dc bit quan trong. Hin nay ngui ta bit duoc
khong 3.500 enzyme khc nhau, nhiu enzyme d duoc
tinh sach, kt tinh v nghin cu cu trc.
3.1.2. Nhng khc bit v dc tnh xc tc, gia xc tc
v co v xc tc enzyme.
Enzyme l cht xc tc sinh hoc, ngoi kh nng xc
tc ging nhu cht xc tc v co bnh thung n cn th
hin mt s tnh cht sau dy:
a) Hiu sut xc tc rt ln.
Su chuyn ho co cht khi s dung emzyme xc tc ln
hon nhiu so vi cht xc tc v co thng thung. V du:
1mol Fe
3+
chi xc tc phn ly duoc 10
-6
mol H
2
O
2
/pht.
Trong khi d mt phn t catalase c mt nguyn t Fe xc
tc phn ly 5.10
6
mol H
2
O
2
/pht;1 gam pepsin trong 2 gi
thu phn duoc 5 kg protein trng luc nhit d bnh
thung. Tuong tu 1gam phn t .-amilase sau 1 giy c th
phn gii 4.000 lin kt glucoside trong phn t tinh bt, cao
hon nhiu so vi xc tc bng cht v co.
b) Tnh dc hiu cao.
Tnh dc hiu cao l mt trong nhng khc bit ch
yu gia xc tc bng enzyme v xc tc bng cc cht
v co khc. Mi enzyme chi xc tc cho su chuyn ho
mt hay mt s cht nht djnh, theo mt kiu phn ng
nht djnh. Dua vo su tc dung c tnh chon loc, ngui ta
c th chia ra mt s kiu dc hiu sau:
- c hiu kiu phn ng. c hiu ny th hin
ch mi enzyme chi xc tc cho mt trong cc kiu phn
ng chuyn ho mt cht nht djnh. V du: phn ng oxy
ho kh, chuyn vj, thu phn, v.v...
- c hiu co cht. L kh nng kt hop ca co cht
vo trung tm hoat dng ca enzyme v bj chuyn ho
dui tc dng ca chng. Dua vo mc d dc hiu ngui
ta lai chia ra mt s kiu nhu: dc hiu tuyt di, l enzyme
chi tc dung trn mt co cht duy nht
3.1.4. Su phn b enzyme trong t bo.
Trong co th enzyme c mt moi m, moi t bo. Nhung
ty theo chc nng ca m m cc t bo trong m thung c
nhng h thng enzyme ring bit.
- Trong nhn c cc enzyme nhu ATP-ase, nucleosidase,
nicotinic-mononucleotide adenylyltransferase, 5
'
-nucleotidase.
- Trong ty lap th c cc enzyme lin quan dn qu trnh
chuyn ho nng luong nhu h enzyme ca chui h hp t bo
v qu trnh phosphoryl ho tao ATP, cc enzyme ca chu trnh
Krebs,v.v...
-Trong lysosome c cha nhiu enzyme thu phn
(hydrolase).
-Trong ribosome c cha cc enzyme cho qu trnh tng
hop protein.
- Trong bo tuong cha nhiu loai enzyme, dc bit c tt
c cc enzyme ca qu trnh dung phn.
- Trn mng t bo cng nhu mng ca nhiu bo quan c
nhng loai enzyme gip cho su vn chuyn mt s cht qua mng
v mt s hoat dng khc ca mng.
.2. Isozyme
3.2.1. c tnh phn t
soyme l cc dang phn t khc nhau ca cng mt
enzyme, xc tc cho mt phn ng sinh ha.
Mc d cng xc tc cho cng mt loai phn ng
sinh ho, nhung do su tn tai cc dang phn t khc
nhau nn c mt s tnh cht l, ho v min djch khc
nhau.
V du: lactatdehydrogengenase (LDH) l enzyme kh
hydrogen thun nghjch gia lactate v pyruvate c khi
luong phn t 130.-140.KDa, cu trc gm 4 tiu don vj
(subunit), cc tiu don vj ny duoc djch m t 2 gene
khc nhau duoc k hiu l H v M (t ch ting Anh H-
Heart v M-Muscle).
Hai loai chui polypeptide d tao nn 5 dang
phn t khc nhau l:
LDH
1
: HHHH
LDH
2
: HHHM
LDH
3
: HHMM
LDH
4
: HMMM
LDH
5
: MMMM
Trong din di theo chiu t m sang duong
th th LDH
1
chay nhanh nht ri dn LDH
2
, LDH
3
,
LDH
4
v LDH
5
chay chm nht.

3.2.2. Vai tr chc nng ca cc isozyme.
T l cc dang isozyme c th thay di tuy theo tui tc,
trang thi sinh l v bnh l. Su tn tai ca nhiu dang
phn t khc nhau ca cng mt enzyme l mt hin tuong
sinh hoc rt quan trong c v mt l lun v thuc tin.
Chng han su phn b khc nhau v LDH trong co th
ca cc loi c xuong sng, loai chui H ch yu tm thy
co tim v loai M ch yu tm thy co xuong. Bi v loai
chui H thung bj c ch bi pyruvat vi nng d qu
tha. Nhung acid pyruvic trong m tim duoc oxy ho hiu
kh mt cch du dn trong ty th d cung cp nng luong
cho m ny, v thng thung khng c su dong ca acid
pyruvic. Tri lai tuy m co xuong c rt nhiu LDH dang M
v m ny c nhiu hoat dng bt thung gy nn nhng
dong pyruvat nht thi, nhung dang M lai khng bj c ch
bi acid pyruvic.
.. Cc enzyme di lp th (allosteric enzyme) v phc h
enzyme (multienzyme).
Trong co th sng cn gp nhng enzyme ngoi
trung tm hoat dng xc tc cn c mt loai trung tm khc
lm nhim vu diu chinh hoat tnh ca enzyme cn goi l
enzyme di lp th.
Tuy trung tm diu chinh v trung tm xc tc c
tc dung h tro nhau, nhung l hai loai cu trc khc bit
nhau.
Trong nhiu trung hop c th "kho trung tm dj
lp th m vn gi duoc hoat dng xc tc ca trung tm
hoat dng. Cc enzyme ny thung duoc cu tao t cc
don vj nh kt hop vi nhau bng cc lin kt yu , nn rt
mm do c th tn tai dui nhiu dang khc nhau v c
th bin di thun nghjch t trang thi ny sang trang thi
khc mt cch d dng.
Ngoi enzyme di lp th cn gp phc h da
enzyme (multienzyme).
l nhng phc hop gm nhiu enzyme
c lin quan dn nhau trong mt qu trnh chuyn
ho nht djnh.
V du phc hop enzyme trong qu trnh tao
acetyl-CoA t acid puruvic duoc goi l phc h
pyruvatedehydrogenase. Phc hop ny gm ba
loai enzyme l pyruvate dehydrogengenase,
dihydrolipoyl transacetylase v dihydrolipoyl -
dehydrogenase.
.4. Nhng quan dim y hc v protein
3.4.1. Protein l nhng phn chc nng ca co th.
Ngoi vai tr l thnh phn chnh trong cu trc ca t
bo v m, protein cn c nhiu chc nng khc quyt djnh
nhng dc dim co bn ca su sng nhu su truyn dat thng tin
di truyn, su chuyn ho cc cht d l cc enzyme, cc khng
th chng lai bnh tt, cc hormon dn truyn cc tn hiu trong
t bo v.v... du c bn cht l cc protein.
3.4.2. Hnh thnh chc nng mi trn co s cu trc protein.
Su pht trin ca sinh hoc phn t dua trn l thuyt trung tm
"DNA F RNA F Protein. Nhu vy, su bin di DNA s dn dn
su bin di cu trc ca phn t protein v do d chc nng
sinh hoc ca n s bj bin di ko theo nhng thay di c lin
quan dn ton b co th.
Trong qu trnh tin ho ca sinh vt su hnh thnh
v thch nghi mt chc nng mi din ra mt giai doan
ljch s lu di.
Su xut hin mt protein mi bin dang thung lun
di km bnh tt.
3.4.3. Su xut hin cc protein bnh l.
Y hoc l ngnh khoa hoc v su sng, ngy nay
vi tin b ca khoa hoc, nhng hiu bit v bnh l
mc d phn t d vuot ra khi gii han ca gii
phu t bo hoc co quan.
Sinh hoc phn t ra di d tao ra cuc cch
mang trong cc quan nim v bnh. T d, su pht
trin ca bnh hoc phn t lun lun di km vi sinh
hoc phn t.
Su bin di cu trc ca mt protein hay su
xut hin cc enzyme c cu trc bt thung du do
yu t di truyn gy nn.
'/ Bnh prion
Prion (Proteinaceous Infectious particle on l
phn t protein v khng cha mt loai acide nucleic
no hoc nu c th cng qu ngn d m ha bt ky
mt protein no m prion c. Trong co th bnh thung
c th c sn prion nhung chng khng gy bnh.
Trong mt diu kin no d prion c th thay di cu
trc v gy bnh. Prion gy nhiu bnh nguy him
dng vt v ngui, gy thoi ha h thn kinh trung
uong v gim st tr tu nhu bnh b din, bnh kuru
ngui. Cc bnh thuc nhm prion duoc phn loai
khoa hoc v CD 10 l A81 v CD-9 l 046.
Tt c bnh do prion duoc bit c nh hung ln
cu trc ca no hoc m thn kinh khc, tt c du
hin khng th diu trj v lun dn dn t vong. Ni
chung, prion duoc hiu theo ngha l mt don vi nhim
v mt l thuyt, song ch gii khoa hoc, th PrP
C
ngh
l mt dang prion protein (PrP) ni sinh, diu ny thy
trong v s m co quan, trong khi PrP
Sc
ngh theo
hung dang gp np ca PrP, chju trch nhim trong
hnh thnh cc mng amyloid v ging ha thn kinh.
Isoforms
Loai protein m prions lm nn (PrP) tm thy khp
co th, trn ngui khe manh v c cc dng vt. Tuy
nhin, PrP tm thy trong cc mu nhim c cu trc khc
nhau v d khng vi cc enzyme protease, cc enzyme
ny trong co th bnh thung c th phn hy cc protein.
Dang protein bnh thung goi l PrP
C
, trong khi
dang protein nhim trng goi l PrP
Sc
ch C ngha l t
bo (cellular) hoc (common) PrP, Sc ngha l 'scrapie',
mt bnh do prion trn cu. Trong khi PrP
C
c cu trc
xc djnh r rng th PrP
Sc
l cc polydisperse chc chn
v xc djnh chua r rng. PrP c th do gp cun nhiu
hay t ln ca dang isoforms d duoc xc djnh r trn in
vitro
PrP
C
PrP
C
l mt protein bnh thung tm thy trn cc mng
t bo. N c 209 amino acids ( ngui), mt cu ni
disulfide, c trong luong phn t 35-36 kDa v ch yu l
cu trc xon alpha. Mt vi dang topological cn tn tai;
mt dang trn b mt t bo hnh m neo qua glycolipid v
2 th xuyn mng
PrP c vai tr quan trong trong qu trnh kt dnh t bo
vi t bo, truyn tn hiu ni bo trong diu kin in vivo,
do d c th lin quan dn truyn tn hiu gia cc t bo
no vi nhau. PrP
Sc
Cc isoform c tnh nhim ca PrP, duoc bit nhu l PrP
Sc
,
c th chuyn thnh dang bnh thung ca PrP
C
proteins
thnh isoform nhim nh vo su thch ng hoc hnh dng;
nguoc lai, su thay di con dung ca cc protein l lin kt
vi nhau.
Mc d cu trc chnh xc 3D ca PrP
Sc
chua bit,
nhung c mt t l cao hon v cu trc -sheet vj tr bnh
thung ca cu trc d-helix
Su kt tp ca cc isoform bt thung ny c cu
trc dang amyloid soi, v t d tch tu thnh tng mng.
iu ny chua r rng nu su kt tp ny gy ra tn thuong
t bo hoc cc phn ng phu dui cc dng ca bnh l
sn c.
Chi c nhng phn t PrP c cu trc sp sp cc
a.a ging ht nhau vi PrP
Sc
mang tnh nhim mi hop
thnh t chc soi pht trin.
Dua theo cc biu hin di truyn ngui ta chia cc protein bnh l
ra lm hai loai ln:
a) Nhng bin di v s luong ca protein:
l su thay di do su tng hoc gim protein no d.
Nhng protein ny vn c cu trc bnh thung v nhu vy
khng c bin di ca gene. M do ri loan qu trnh diu ho
sinh tng hop protein
Trong trung hop l protein enzyme th nhng lch lac v
s luong enzyme s dn dn nhng ri loan dy chuyn chuyn
ho.
b) Nhng bin di v cht luong protein:
l nhng ri loan v cu trc protein do gene bi bin
di, dn dn cu trc protein thay di ko theo su thay di chc
nng sinh hoc ca protein d. V du, su bin di cu trc ca
hemoglobin (Hb) l protein c chc nng vn chuyn oxygen
trong mu dn dn bnh thiu mu, hay nhu bnh thiu mu do
hng cu hnh lui lim...
3.4.4. Cu trc v chc nng ca protein min djch.
Tham gia vo h thng min djch c nhiu co
quan, nhiu loai t bo v dc bit nhiu loai protein
thuc hin cc chc nng ring bit tao nn hiu qu
min djch dc hiu v khng dc hiu. Cc protein
min duoc nhc dn nhiu hon c l cc khng th,
b th v cc cytokine.
a) Cc khng th (antibody).
Tt c cc phn t khng th d duoc chng minh l
cc globulin c chc nng min djch (vit tt l g:
mmunoglobulin) v c bn cht l glycoprotein. Cc
khng th duoc chia thnh 5 lp. Tuy theo cu trc v
chc nng min djch l gG, gA, gM, gD, gE.
Cu trc ca phn t khng th duoc hnh thnh
t hai loai chui polypeptide l chui nng (k hiu:
H=Heavy chain) c khi luong phn t t 53-59 kDa v
chui nhe (k hiu:L=Light chain) c khi luong phn t
22-26 kDa.
C bn chui duoc gn vi nhau bng cu
disunfid (S-S). Trung tm hoat dng l phn lin kit vi
khng nguyn (hnh 1.2) nm vng tn cng N ca
chui nng v chui nhe c cu trc chi khong 8-10
amino acid.
H Knh 1.2 Cu trc chung cua phn tu khng th (Ig)
b) B th (complement).
L nhng protein huyt tuong phn ng vi nhau
nhm tn cng cc dang tc nhn gy bnh.
H thng b th bao gm khong 40 protein c
chc nng dp ng min djch, chng vi sinh vt v dp
ng vim.
V chc nng, cc khng th tuong tc dc hiu
vi tc nhn truyn nhim bnh, cn h thng b th
duoc c djnh ln tt c cc khng th d thuc hin chc
nng min djch.
Cc thnh phn ca b th tuong tc gia chng
vi nhau v cc yu t khc ca h thng min djch.
c) Cc cytokine.
L ton b cc phn t duoc tit ra bi cc t bo ca
h thng min djch, tham gia vo hoat dng tn hiu gia cc
t bo trong hoat dng dp ng min djch. Tt c cc
cytokine du c bn cht protein hay glycoprotein v duoc
phn loai nhu sau:
- Cc interferons (FN): c cc dang --FN, .-FN v -
FN, c chc nng ngn nga ca mt s virus gy bnh.
- Cc interleukin (L): c cc dang t L1 dn L 13,
chng c nhiu chc nng, nhung ch yu l kim tra su bit
ho v sinh sn t bo.
- Cc yu t kch thch qun lac (CSF): c chc nng
kim tra su phn chia v sinh sn ca cc t bo ngun v
cc t bo mu so khai.
- Cc cht dn truyn sinh hoc (mediator): l nhng
protein ca giai doai dp ng min djch cp tnh.
W 3.4.5. Cu trc v chc nng ca protein vn chuyn.
W Trong co th c nhng protein lm nhim vu vn chuyn nhu
hemoglobin, mioglobin, hemocianin vn chuyn O
2,
CO
2
v H
+
di
khp cc m, cc co quan trong co th. Ngoi ra cn c nhiu
protein khc nhu lipoprotein vn chuyn lipid, ceruloplasmin vn
chuyn dng (Cu) trong mu v.v...Mt trong nhng protein lm
nhim vu vn chuyn duoc nhc dn nhiu nht d l hemoglobin.
Phn t duoc cu tao t bn tiu don vj (subunit), hai tiu don vj -
v hai tiu don vj ..
W
W HKnh 1.3 Cu trc ca phn t hemogIobin
W Mi tiu don vj ni vi mt heme bng lin kt khng
phi cng ho trj. Khi so snh vi protein cng chc
nng nhu myoglobin co v leghemoglobin thuc vt l
nhng protein c cu trc chi mt tiu don vj (monomer)
thy rng cc tiu don vj cu trc kh ging nhau
3.4.6. Cu trc chc nng v vai tr ca lectin.
Lectin l nhng protein hay glycoprotein khng phi
ngun gc min djch, lectin c kh nng ngung kt vi nhiu
loai t bo, cng nhu nhiu loai dung hoc cc hop cht
cha dung c tnh cht chon loc.
Hu ht lectin c cu trc bc 4, vi khi luong phn t
giao dng trong pham vi kh rng t hng ngn cho dn hng
trm ngn Dalton. V du: lectin t r cy &rtica dioica (ho gai
&rticaceae) c Mr=8,5 KDa trong khi d loi sam bin chu
(Tachypleus tridentatus) c Mr=700.KDa.
V chc nng, ngui ta thy rng mc d lectin khng
phi l khng th chng lai tc nhn gy bnh nhung chng c
vai tr bo v co th nh tuong tc vi mng t bo v gy
ngung kt t bo ca chng.
3.4.7. Nhng chc nng khc ca protein
Trong co th ngoi cc protein dm nhn chc
nng xc tc nhu enzyme, chc nng vn chuyn nhu
hemoglobin, mioglobin, lipoprotein, v chc nng bo
v nhu cc khng th min djch, cc protein dc t
nhu enzyme noc rn, lectin v.v..., protein cn tham gia
nhiu chc nng quan trong khc nhu:
- Cc protein lm nhim vu kch thch diu ho
qu trnh trao di cht nhu cc hormon
- Cc protein lm nhim vu cu trc nhu v
virus, mng t bo, colagen da, fibrolin to
- Cc protein lm nhim vu co rt nhu myosin,
actin soi co
- Cc protein lm nhim vu du tr nhu casein
ca sa, ovalbumin ca trng, v.v...