HOA HOC PROTID

MUC TIEU BAI HOC

Viet cong thc cua 20 acid amin thng gap
trong phan t protein
Trnh bay c cac tnh chat cua acid amin: tnh
tch ien, tnh chat vat ly va hoa hoc
Trnh bay c cac tnh chat cua acid amin: tnh
tch ien, tnh chat vat ly va hoa hoc
Neu c ten mot so peptid co chc nang sinh
hoc
Trnh bay c cac lien ket va cac bac cau truc
cua protein
Trnh bay c cac tnh chat cua protein: khuech
tan, tch ien, hoa tan va ket tua, bien tnh.
PROTEIN LA G?
Protein la ai phan t polymer do cac phan t
acid amin (monomer) noi vi nhau bang lien
ket peptid. Trnh t cua cac acid amin do gen ket peptid. Trnh t cua cac acid amin do gen
quyet nh.
Protein co nhieu chc nang the hien trong
cac vai tro: cau truc, xuc tac, van chuyen,
van ong, dinh dng va d tr, bao ve, ieu
hoa hoat ong cua c the.
PROTEIN
Acid amin (o-amino acid)
Peptid Peptid
Protein
PROTID
ACID AMIN Cau truc
Acid amin
* mot nhom amin (-NH
2
)
* mot nhom carboxyl (-COOH) * mot nhom carboxyl (-COOH)
cung gan vao carbon o
* mot chuoi ben (-R).
Acid amin c ve
* - N ben trai
* - C=O ben phai
Goc R
ACID AMIN Cau truc
o-Amino acid
ACID AMIN Cau truc
Trong thien nhien co khoang 300 AA
Trong protein/sinh vat co 20 AA
(o la cac o-amino acid)
Mot so AA khong phai la o-amino acid:
|-alanin, -aminobutyric acid
ACID AMIN ong phan
Tr glycin, tat ca AA
khac eu co carbon khac eu co carbon
bat oi
(Carbon o nam v tr
trung tam bat oi )
ACID AMIN ong phan
Trong protein ch cha L-o-amino acid
ACID AMIN ong phan
D, L da vao D- va L- glyceraldehyd
(L: levorotatory; D: dextrorotatory)
ACID AMIN Phan loai
Da vao chuoi ben (-R)
Khong phan cc
Phan cc va khong tch ien
Phan cc va tch ien
ACID AMIN Phan loai
AA khong phan cc hydrophobic water fearing
ACID AMIN Phan loai
AA khong phan cc hydrophobic water fearing
ACID AMIN Phan loai
AA phan cc khong tch ien
(chuoi ben cha cac nhom phan cc kho ion hoa)
ACID AMIN Phan loai
AA phan cc khong tch ien
(chuoi ben cha cac nhom phan cc kho ion hoa)
ACID AMIN Phan loai
AA phan cc va tch ien am pH c the
(chuoi ben cha nhom carboxyl)
ACID AMIN Phan loai
AA phan cc, tch ien dng pH c the
(chuoi ben cha nhom amin)
ACID AMIN Phan loai
Mot so AA ac biet
Hydroxylysine, hydroxyproline - collagen
-Carboxyglutamate - prothrombin
3-Monoiodotyrosine, 3,5-diiodotyrosine, T
3
, T
4
-
hormon giap trang va cac tien chat
Citrullin, ornithin - sinh tong hp ure
ACID AMIN Phan loai
Vai tro cua (R) / AA trong chuoi polypeptid
Vong imidazole (His, trong Hb, Mb)
Chc OH (Ser, Thr), SH (Cys) Chc OH (Ser, Thr), SH (Cys)
Cysteine-SH + HS-Cysteine <-----> Cysteine-S-S-Cysteine
COO
-
, NH
3
+
cua (R) cac AA acid hay base
ong vai tro quan trong trong hnh thanh cau truc, tham
gia cac nhom trung tam hoat ong cua enzym
ACID AMIN Phan loai
Vai tro cua cac AA trong chuoi polypeptid
AA k nc khong tng tac trong moi trng
nc, chiem phan ln be mat pha trong phan t nc, chiem phan ln be mat pha trong phan t
protein. Loai nay khong b ion hoa, khong tao LK
hydro.
AA a nc tng tac vi moi trng nc, tao
LK hydro vi nc va chiem t le ln tren be
mat ngoai cua protein, hay co mat cac trung tam
hoat ong cua enzym.
ACID AMIN ien tch
pH trung tnh, AA dang lng cc
ACID AMIN Tnh chat
pH trung tnh (pH mau: 7,4; pH noi bao: 7,1)
Dipolar (zwitterion)
ACID AMIN ien tch
R-COOH R-COO
-
+ H
+
R-NH
+
R-NH + H
+
R-NH
3
+
R-NH
2
+ H
+
Theo quan niem Bronsted th AA cha t
nhat 2 nhom acid phan ly yeu
AA la acid yeu co the phan ly nhieu lan
ACID AMIN ien tch
ACID AMIN Tnh chat
Dang ion hoa cua AA thay oi tuy theo pH moi
trng thay oi [C] cua cac dang ion
Tai pH moi trng:
o A
+/-
= MAX
o A
-
= A
+
= MIN
o E IEN TCH = 0
AA khong di chuyen trong ien trng
pH
i
(isoelectric)
(pH ang ien)
ACID AMIN Tnh chat
Tai pH moi trng < pHi
AA hoat ong nh base
R-CH-COO
-
R-CH-COOH R-CH-COO
-
R-CH-COOH
NH
3
+
NH
3
+
Ton tai ca 3 dang ion A
+/-
, A
-
, A
+
Cation A
+
chiem t le nhieu nhat
Trong ien trng, AA chuyen ve cc (-)
H
+
ACID AMIN Tnh chat
Tai pH moi trng > pHi
AA hoat ong nh acid
R-CH-COO
-
R-CH-COO
-
R-CH-COO
-
R-CH-COO
-
NH
3
+
NH
2
Ton tai ca 3 dang ion A
+/-
, A
-
, A
+
Anion A
-
chiem t le nhieu nhat
Trong ien trng, AA chuyen ve cc (+)
H
+
ng chuan o glycin 0,1M, 25
o
C
pK1 - COOH
pK2 - NH
3
+
ACID AMIN Tnh pI
ACID AMIN Tnh chat
Acid aspartic
A
+
A
+/-
A
+/-/-
A
-/-
pK pK pK pK
1
pK
2
pK
3
Arginin
A
+/+
A
+/+/-
A
+/-
A
-
pK
1
pK
2
pK
3
ACID AMIN Tnh chat
Tnh chat vat ly
De tan trong dung moi phan cc
V ngot kieu ng (natri glutamat: v V ngot kieu ng (natri glutamat: v
ngot kieu am)
Pho hap thu:
Tnh chat vat ly
Cac acid amin khong hap thu anh sang nhn
thay c (khong mau), va (tr acid amin thm thay c (khong mau), va (tr acid amin thm
Phe, Tyr, His va Trp) hap thu anh sang vung
tia hong ngoai
Ch Phe, Tyr, His va Trp hap thu vung UV
o nong o AA tot nhat 280 nm
ng dung: dd acid amin, nc mam
Tnh chat vat ly
Tnh chat hoa hoc
PHAN NG DO NHOM o-NH
2
VA o-COOH
1. Phan ng NINHYDRIN 1. Phan ng NINHYDRIN
2. PHAN NG TAO LIEN KET PEPTID
PEPTID
Ten: Acid glutamic - Glycin - Alanin - Lysin
K hieu ba ch: Glu-Gly-Ala-Lys
K hieu mot ch: EGAK
PEPTID - Cau tao
PEPTID
C
O
N C N
H
Lien ket
PEPTID
AA1 AA2 AA3 AA4
(Ban chat la LK amid)
PEPTID
Vi n AA, co n-1 lien ket peptid
Co 2 au: N tan va C tan
N tan pha ben trai - au chuoi
C tan pha ben phai - cuoi chuoi
Ten goi theo ten rieng
Glutathion, glucagon, insulin,
ACTH
PEPTID- tnh chat
Phan t peptid co:
Nhom o-amin t do au N tan
Nhom o-carboxyl t do au C tan
Bo khung: mach lien ket peptid
Chuoi ben: cac goc R cua cac AA
Co tnh chat lng tnh giong AA
PEPTID- tnh chat
Moi phan t peptid co mot pH
i
tng ng
pH moi trng > pH
i
: Peptid tch ien (-)
pH moi trng < pH
i
: Peptid tch ien (+)
Phan ng nhan biet lien ket peptid (Biuret)
OH
-
, Cu
++
Phc tm hong Biure
(-CO-NH- )
PEPTID- tnh chat
-NH
3
OH
-
Cu
++
Phc tm hong Biure
PEPTID- tnh chat
Tng t vi protein
Peptid co t 3 AA (2 lien ket peptid) th phan ng biuret (+)
PEPTID co hoat tnh sinh hoc
Neuropeptid: Co mat nao bo, anh hng len
hoat ong cua TKTW, chu yeu do tuyen yen va vung
di oi.
Enkephalin (5AA), endorphin (15AA); oxytocin
Hormon peptid:
Insulin (51AA); glucagon (29AA); gastrin (16AA)
Peptid khang sinh: do vi khuan, nam tao ra; cha ca
L va D AA; cha mot so AA khong co trong protein
PEPTID co hoat tnh sinh hoc
Gramicidin S: Val-Orn-Leu-D.Phe-Pro
Pro-D.Phe-Leu-Orn-Val
tac dung tren vi khuan Gram (+), lam h mang tac dung tren vi khuan Gram (+), lam h mang
phospholipid cua vi khuan.
Peptid tham gia he thong oxy hoa-kh
Glutathion: glutamyl-cystein-glycin
2 G-SH 2 G-S-S-G
-2H
+2H
PEPTID co hoat tnh sinh hoc
PROTEIN: Phan loai
Nhieu he thong phan loai do s phc tap ve
Cau truc - Tnh chat ly hoa Chc nang sinh hoc
Da vao tnh hoa tan: Alb (H
2
O); globulin (muoi loang) Da vao tnh hoa tan: Alb (H
2
O); globulin (muoi loang)
Da vao hnh dang: Protein si (collagen, elastin);
Protein cau (enzym, alb, myoglobin)
Da vao chc nang: Cau truc, van chuyen, xuc tac
Da vao tnh chat vat ly: phan biet cac protein gan nhau
LP (chylomicron, VLDL, LDL, HDL)
PROTEIN: Phan loai
PROTEIN: Phan loai
Protein huyet thanh: Albumin, globulin (o1,o2,|,)
Da vao tnh chat hoa hoc :
Protein thuan: ch cha cac AA Protein thuan: ch cha cac AA
Albumin: pHi 4,6-4,7 MW 35.000-70.000
Globulin: pHi 5,2-6,8 MW 90.000-150.000
Histon: pHi 9-11, cha nhieu AA base
Keratin: MW >2trieu Cha nhieu cystin
Collagen: thuy phan tao gelatin; MW 350.000
PROTEIN: Phan loai
Protein tap (protein thuan + nhom ngoai)
Nucleoprotein
Cromoprotein Cromoprotein
Lipoprotein
Glycoprotein
PROTEIN-Lien ket
Ban chat la LK amid (LK peptid hi ngan hn LK n khac).
Tnh chat LK oi mot phan, han che quay quanh LK
Lien ket peptid
peptid, nen 4 nguyen t CO-NH- nam tren cung mot
mat phang
PROTEIN-Lien ket peptid
LK peptid khong quay
Lien ket u (-N-Co) (Phi) Lien ket u (-N-Co) (Phi)
va (-CO-Co -) (Psi) quay
t do
PROTEIN-Lien ket peptid
Chuoi polypeptid c coi la co nhieu mat
phang, co 2 goc quay gia moi mat phang
PROTEIN-Lien ket peptid
V anh hng khong gian, Phi va Psi quay han che tao
cau hnh Trans > Cis tao cau truc khong gian protein
PROTEIN-Lien ket disulfid
LK ong hoa tr gia 2 cystein tren 1 chuoi hay 2
chuoi khac nhau.
ong vai tro quan trong trong duy tr cau truc bac III ong vai tro quan trong trong duy tr cau truc bac III
Khi muon phan tch protein phai pha v LK nay
cystein cystein
S S
cystein
cystein
S
S
PROTEIN-Lien ket Hydro
La lc hut tnh ien gia H tha ien tch (+) va O
(hay N, Cl) tha ien tch (-) tren cung chuoi hay cac
chuoi khac nhau. Trong protein, so lng LK hydro ln.
Quan trong trong hnh thanh va duy tr cau truc bac II
0xy
H
Quan trong trong hnh thanh va duy tr cau truc bac II
PROTEIN-Lien ket ion
La lc hut tnh ien gia nhom COO
-
(Glu, Asp)
vi NH
3
+
(Lys, Arg), con goi la LK muoi
Glu
Lys
COO
-
NH
3
+
PROTEIN-
Tng tac k nc gia cac R
Tng tac gia cac goc khong phan cc ben trong
protein (lien ket Van der WAAls): yeu, khong la lien
ket thc s; quan trong trong duy tr cau truc protein. ket thc s; quan trong trong duy tr cau truc protein.
alanin
valin
R
R
PROTEIN-Lc Van de Waals
La lc tng tac gia cac nguyen t, phan t
khoang cach ngan
cc ky yeu
PROTEIN-Vai tro cua cac LK
Cac AA noi vi nhau bang lien ket peptid tao nen
chuoi polypeptid.
Trnh t cac AA la mot dang thong tin quyet nh
cau truc cua protein. cau truc cua protein.
Cac tng tac khong hoa tr (thuoc goc R) ong
vai tro quan trong trong cau truc cua protein.
Cac AA lien ket vi nhau bang cac lien ket co o
linh hoat gii han e tao nen cau truc ong hoc 3
chieu cua phan t protein.
Chnh cau truc se quyet nh chc nang cua protein
PROTEIN-cau truc
Vao nhng nam 1950, Linus Pauling at ten cho
nhng cau truc ong tm ra nh nhieu xa tia X la xoan
alpha, la gap beta. alpha, la gap beta.
Nay them mot so cau truc khac nh uon beta, cau
truc collagen
PROTEIN-cau truc bac 1
Bieu th trnh t cac AA trong chuoi polypeptid
V tr cua cac cystein kha nang tao LK disulfid
PROTEIN-cau truc bac 2
Bieu th s tng quan cua cac AA gan nhau trong
chuoi polypeptid
Lien ket hydro ong vai tro quan trong Lien ket hydro ong vai tro quan trong
Ba dang cau truc bac 2 thng hay gap
Xoan o (Alpha helix)
La gap | (Beta-Pleated Sheet)
Cau truc uon | (| turns)
PROTEIN- xoan o
Cau truc hnh roi, si polypeptid xoan quanh truc
phan t tao phan loi, goc R cua AA nho ra ngoai
Ben vng nh LK hydro, moi vong xoan co 3,6 AA Ben vng nh LK hydro, moi vong xoan co 3,6 AA
Protein nhieu xoan o
(Hb, Mb)
Protein t xoan o
(Chymotrypsin)
PROTEIN-xoan o
PROTEIN- xoan o
PROTEIN- La gap |
Chuoi polypeptid trai ra co hnh zigzag
Chuoi polypeptid ke ben cung chieu: song song Chuoi polypeptid ke ben cung chieu: song song
Chuoi polypeptid ke ben khac chieu: oi song
C cau ben la do lien ket hydro gia cac chuoi
peptid bac I xa nhau
PROTEIN- La gap |
i song
Song song
PROTEIN- La gap |
PROTEIN- La gap |
A v B: i song; B v C: song song
PROTEIN- La gap |
PROTEIN-Cau truc bac 2 khac
Cau truc uon |: cho phep xon o va la gap | sap xep
side-by-side (prolin va glycin)
bab
PROTEIN-Cau truc bac 2 khac
Xoan collagen
cha nhieu glycin, prolin
va hydroxyprolin tao
dang ba si gn vi nhau dang ba si gn vi nhau
nh LK hydro.
PROTEIN-cau truc bac 3
Cau truc khong gian 3 chieu trong 1 chuoi PP.
Co ac iem:
Co cac cau truc bac 2 giong hay khac nhau Co cac cau truc bac 2 giong hay khac nhau
Cuon lai tao thanh khoi ac chac
LK disulfid ong vai tro quan trong trong
duy tr cau truc nay
PROTEIN-cau truc bac 3
PROTEIN-cau truc bac 4
Co t 2 chuoi polypeptid bac 3 tr len, tao thanh
protein oligomer. Tng chuoi PP c goi la
protomer, monomer, di (ban, tieu) n v. protomer, monomer, di (ban, tieu) n v.
Cac chuoi PP lien ket vi nhau qua cac lc
khong ong hoa tr: lien ket H va lien ket tnh
ien.
MW cua hau het cac protein bac 4 >50.000 Da.
PROTEIN-cau truc bac 4
Hemoglobin (Hb) gom 4 tieu n v:
2 chuoi o, 2 chuoi |
PROTEIN
Ch thay oi 1 AA trong phan t protein dan ti thay
oi hoat tnh hay chc nang cua protein o.
Th du: Thieu mau hong cau liem (Sickle cell anemia) Th du: Thieu mau hong cau liem (Sickle cell anemia)
HbA: X-X-X-X-X-Glu (chuoi |)
HbS: X-X-X-X-X-Val (chuoi |)
lam thay oi hnh dang cua hong cau t hnh da thanh
hnh liem, giam tnh an hoi va tang kha nang v hong
cau
PROTEIN
HC hnh liem
PROTEIN-Tnh chat
Ly tnh
Protein/nc tao dd keo (1<d<100nm)
Khuech tan trong dd cham Khuech tan trong dd cham
Khong qua c mang
ban tham
Tham tch loai muoi
PROTEIN-Tnh chat
Ap dung:
Than nhan tao Than nhan tao
Tham phan phuc mac
PROTEIN-Tnh chat
Protein/nc tao ap suat (p) tham thau goi p keo
P keo <30-40 mmHg nhng quan trong trong van
chuyen nc, chat dinh dng va san pham chuyen nc, chat dinh dng va san pham
chuyen hoa qua thanh mach
Mao M
Mao TM
Mao mach
ASTT> ASK
ASTT < ASK ASTT = ASK
40 28
16 40
90%
10%
Bach huyet
PROTEIN-Tnh chat
Tnh chat acid-base
Giong AA va peptid, protein/ddch ton tai ca 3
dang ion P
+/-
, P
+
, P
-
. Tuy pH moi trng so vi pH
i
dang ion P , P , P . Tuy pH moi trng so vi pH
i
cua protein ma protein tch ien (+) hay (-)
ng dung ien di protein (electrophoresis)
NT: da tnh chat tch ien pH mt = pHi
di chuyen trong ien trng
ien tch
Trong lng
Kch thc
PROTEIN-Tnh chat
ien di o
Protein
huyet thanh
(serum)
Alb: pHi 4,6 5,2
Globulin: pHi 5,2- 6,8
PROTEIN-Tnh chat
ien di o
PROTEIN-Tnh chat
ien di o
PROTEIN-Tnh chat
Hoa tan va ket tua
Dd keo ben vng nh
Cac yeu to anh hng o hoa tan
Lp ao nc (hydrat hoa)
S tch ien cung dau
Cac yeu to anh hng o hoa tan
pH
Nong o muoi : muoi trung tnh
Nong o muoi thap: tang o tan (salting in)
Nong o muoi cao: giam o tan, co the gay
tua protein (salting out)
pH = pH
i
: o tan thap nhat
pH = pH
i
: o tan gia tang
PROTEIN-Tnh chat
ng dung
Phng phap diem tch: loai muoi khoi dd protein
Tua globulin: muoi amoni sulfat ban bao hoa
Dung moi: Alcol, ceton, amoni sulfat giam o tan
Nhiet o: 0
o
C en 40
o
C: o tan tang
>40
o
C: o tan giam, mat tnh ben vng
Tua globulin: muoi amoni sulfat ban bao hoa
Tua albumin: muoi amoni sulfat bao hoa
PROTEIN-Tnh chat
S bien tnh cua protein
Protein
Bien tnh
T
o
, pH, chat tay
Acid manh, kim loai nang Acid manh, kim loai nang
Cau truc b ao lon
Ch con lien ket peptid
Goc k nc (R) quay ra ngoai
Tnh chat ban au b mat
o tan giam, mat tnh sinh khang nguyen
Mat hoat tnh sinh hoc, de tieu hoa
PROTEIN
-Tnh chat
S bien tnh thuan nghch
Trong mot so ieu kien nhat
nh, protein b bien tnh co the
tr lai dang ban au vi cau
nh, protein b bien tnh co the
tr lai dang ban au vi cau
truc, tnh chat va chc nang
nguyen thuy
Th du: ribonuclease
PROTEIN-Tnh chat
S bien tnh khong thuan nghch
khong tr lai trang thai ban au
Th du: Long trang trng luoc Th du: Long trang trng luoc
ng dung
Chiet xuat, tinh che protein (enzym) e
phong s bien tnh (thao tac t
o
lanh, am bao
pH)