HA HQC HA HQC

PORPHYRIN V PORPHYRIN V
HEMOGLOBIN HEMOGLOBIN
BS Trn Ngc Minh BS Trn Ngc Minh
B mn Ha Sinh B mn Ha Sinh- -khoa Y khoa Y
Muc tiu Muc tiu
Phn loai cromoprotein.
M t cu tao ca HEM (v hnh).
Phn tch cu tao ca GLOBN.
Phn tch su kt hop ca HEM v GLOBN.
Hiu vai tr ca Hb trong vic vn chuyn cc kh.
Phn tch su kt hop thun nghjch ca HEMOGLOBN (Hb)
vi oxy (O
2
) v carbon dioxyd (CO
2
).
M t tnh cht kt hop ca Hb vi oxyd carbon (CO).
Nu tn 3 cht oxy ha Hb thnh Met-Hb.
Nu tnh cht enzym ca Hb.
Ni Dung Ni Dung
. A CUONG
1.1. Cromoprotein.
1.2. Porphyrin.
. HEMOGLOBN
2.1. HEM.
2.2. GLOBN.
2.3. Su kt hop HEM v GLOBN.
. TNH CHT CA HEMOGLOBN
3.1. Kt hop thun nghjch vi oxy (O
2
).
3.2. Kt hop thun nghjch vi carbon dioxyd (CO
2
).
3.3. Kt hop vi oxyd carbon (CO).
3.4. Oxy ha Hb tao methemoglobin (MetHb).
3.5. Tnh cht enzym ca Hemoglobin
W Chc nng chnh ca hng cu:
W 'n chuyn O2 t phi dn m
W 'n chuyn CO2 t m dn phi
W Chc nng ny thuc hin duoc nh hng
cu c hemoglobin (Hb)
W Mi hng cu c 640 triu phn t Hb
Gii thiu Gii thiu
0rythrocyt0
ai cuong ai cuong
W Cromoprotein: protein tap, nhm ngoai l
cht mu
W Hai loai:
Porphyrinoprotein: nhm ngoai l nhn
porphyrin
Cromoprotein c nhm ngoai khng
phi l nhn porphyrin
Cromoprotein (1) Cromoprotein (1)
Porphyrinoprotein: c nhm ngoai l nhn
porphyrin
Th du :
O Hb (hemoglobin): sc t d ca hng
cu.
O Myoglobin: Sc t h hp trong t bo
co dng vt c xuong sng.
O Mt s oxydoreductase: cc enzym xc
tc phn ng oxy ha kh nhu cytocrom.
Brenner, S.E. Nature Reviews Genetics. 2001. 2, 801-809
Myoglobin v Hemoglobin
Myoglobin
Hemoglobin
Cromoprotein (2) Cromoprotein (2)
Cromoprotein cha nhm ngoai khng c
nhn porphyrin.
Th du:
O Flavoprotein F cha riboflavin
O Ferritin F cha st
O Hemocyamin F cha dng
Porphyrin (1) Porphyrin (1)
W Cu tao ha hoc ca porphyrin bt ngun t
vng pyrol.
W Pyrol :
N
CH
CH C H
C H
H
N
H
Porphyrin (2) Porphyrin (2)
W Porphin: bn vng pyrol lin kt vi nhau qua 4
cu ni methenyl (-CH=)
5 6
N H
N
NH
N
1 2
3
4 7
8
1
2
3
4
5 6
7
8

'
(1) (2)
CH C
C CH
CH
N H
CH C
C
NH
N
C C H
C H
C H C
C H C
N
C H CH
C
C H CH
(3)
(4)
(5) (6)
(7)
(8)

'

o
Porphyrin (3) Porphyrin (3)
W Porphyrin: porphin + gc ha hoc
(tai cc vj tr 1, 2, 3, 4, 5, 6, 7, 8)
Ty theo thnh phn cc nhm th m ta
c cc loai porphyrin khc nhau :
protoporphyrin (4M, 2', 2P);
coproporphyrin (4M, 4P);
uroporphyrin (4A, 4P).
Cc gc th ca porphyrin Cc gc th ca porphyrin
Tn gc ha hoc Cng thc 'it tt
Methyl
Ethyl
Hydroxyethyl
'inyl
Gc a. acetic
Gc a. propionic
-CH
3
-CH
2
-CH
3
-CH
2
-CH
2
OH
-CH=CH
2
-CH
2
-COOH
-CH
2
-CH
2
-COOH
M
E
E-OH
'
A
P
Mt s porphyrin Mt s porphyrin
M '
M
'
M P
P
M
M P
M
P
M P
P
M
A P
A
P
A P
A
P
M P
M
P
M P
M
P
A P
A
P
A P
P
A
M E
M
E
M P
P
M
OH
OH
Protoporphyrin X Uroporphyrin Uroporphyrin
Coproporphyrin Coproporphyrin Hematoporphyrin
Porphyrin ( Porphyrin (4 4) )
W Tnh cht:
Hop cht c mu.
Base yu.
Acid.
im dng din (pHi) 3 - 4,5.
Tnh tan phu thuc s nhm carboxyl th
Hemoglobin (Hb) Hemoglobin (Hb)
Hb mt cromoprotein, hay chnh xc hon l mt
porphyrinoprotein.
Cu tao Hb gm 2 phn:
hem : nhm ngoai
globin : protein
Hem: protoporphyrin X + Fe
+2
Hng cu ngui cha khong 32% Hb ( 15g/100ml mu).
Ty nhm GLOBN, khi kt hop vi HEM s tao thnh nhng
loai Hb khc nhau.
Hem Hem
L protoporphyrin X gn vi 1 nguyn t Fe
+2
bng 2 ni cng ha trj v 2 ni phi tr.
Fe
+2
HEM porphin: 4 vng pyrol
Protoporphyrin
8 nhm th
(M, ', M, ', M, P, P, M)
Hem Hem
N
Fe N N
M '
M
'
M P
P
M
N
Hem Hem
Khi Hem bj oxy ha:
HEM HEMATN HEMN (tm)
Fe

Fe

(tinh th Teichman)
Fe
N
N
N
N
OH
Fe
N
N
N
N
Cl
Globin Globin
O Cu trc bc I:
Chui H
2
N COOH (141 aa)
H
2
N COOH (146 aa)
O Cu trc bc II:
70% cc aa chui & tao thnh nhiu doan xon
A B C D
A B B C C D
O Cu trc bc III: cc chui & gp khc.
B
oan khng xon
A A
NH
2
Cu trc bc hai, bc ba ca globin vi cc vng xon alpha
'ng xon alpha
www.people.virginia.edu/ ~rjh9u/giI/Iig17mod.giI
http://www.biochem.ucl.ac.uk/bsm/cathnew/
Globin Globin
Goi tn cc aa trn chui polypeptid bng th tu
ca chng trn doan xon
Td: Asp G1
His E7; His F8
xc djnh su khc bit gia cc Hb, ngui ta phi
xc djnh th tu cc aa trong cu trc bc ca
Globin.
HbA & HbS: trn chui : Glu (6) HbA bj th bi 'al (HbS)
HbA & HbC: trn chui : Glu (6) HbA bj th bi Lys (HbC)
F Pauling coi d l biu hin ca bnh l phn
t.
Globin Globin
Mu tu A, B, C
K hiu Hb:
noi tm ra.
1961, Gerand & ngram: k hiu theo chui
polypeptid
Td: HbA:
2
A

2
A
HbS:
2
A

2
S

2
A

2
6gluFval
HbC:
2
A

2
C

2
A

2
6gluFlys
Sickle Cell Mutation
+O2
-O2
+O2
-O2
5'
l 2
3'
Chromosome 16
5' 3'
Chromosome 11 s o
G A


CCT GAG GAG
-Pro-Glu-Glu-
5 6 7
CCT GTG GAG
-Pro-'al-Glu-
5 6 7

S
Normal (HbA) Abnormal (HbS)

0XY-8TATE 0E0XY-8TATE
$ kt hp gia Hem v Globin $ kt hp gia Hem v Globin
Mi chui polypeptid ca GLOBIN kt hp vi mt HEM
qua 2 lin kt phi tr gia Fe
+2
v N ca nhn Imidazol
ca aa histidin E7 & F8 tao thnh 1 bn don v ca phn t
Hb.
(His xa)
N
N
Fe
N
N
HS (E7)
HS (F8)
N
N
Fe
N
N
O=O
HS (F8)
$ kt hp gia Hem v Globin $ kt hp gia Hem v Globin
Nguyn t Fe gm
6 lin kt:
+ 2 lin kt cng ha tr
+ 4 lin kt phi tr
Khi kt hp oxy,
nguyn t Fe vn
ha tr 2 F dy l
phn ng gn
oxy ch khng
phi l s oxy ha.
N
N Fe N
N
O
2
N
HS HS
N
N Fe N
N
N N
HS HS
N
(E7)
Bn don v Hb t do v kt hp oxy Bn don v Hb t do v kt hp oxy
GLOBN
M
'
M
'
M P
P
M
Fe
N
N
N N
N
O
2
GLOBN
M
'
M
'
M P
P
M
Fe
N
N
N
N N
N
N
Hemoglobin Hemoglobin
Protein hnh cu, dung knh 5,5 nm.
Gm 4 bn don v ging nhau tng di
mt.
Mi bn don v:
1 Hem v 1 chui polypeptid.
Trong lung phn t Hb: 64456 Da.
Khong cch gia cc Hem/Hb 2,5nm.
Fe
+2
trong Hb 0,34%.
Bn don v ca Hb Bn don v ca Hb
Phn t Hb hon chnh Phn t Hb hon chnh
g|oo|r g|oo|r
g|oo|r g|oo|r
Hb A tetramer
Tng hp Hemoglobin Tng hp Hemoglobin
Cc chui alpha & beta Cc chui alpha & beta
Tng hp globin Tng hp globin
- Nhng loai globin khc nhau kt hp
vi hem d tao thnh cc Hb khc
nhau
- Nhng gen m ha cc chui globin:
cluster: gen , , o v s nm trn nhnh
ngn nhim sc th 11
cluster: gen v nm trn nhnh ngn
nhim sc th 16
Globin bt du duc tng hp t tun
th 3 ca thai ky
- Phi
Hemoglobin Gower I (
2
s
2
)
Hemoglobin Portland (
2

2
)
Hemoglobin Gower II (
2
s2)
- Thai : HbF (
2

2
), HbA (
2

2
)
- Ngui ln : HbA, HbA
2
(
2
o
2
), HbF.
Tng hp globin Tng hp globin
Biu d qu trnh tng hp Biu d qu trnh tng hp
cc chui globin cc chui globin
Gan Lch Ty xuong
Ti non hong
sinh
Sau sinh(tun)
Tui thai(tun)
%
Hb A Hb A
2
Hb F
Cu trc
2

2

2
o
2

2

2
% 95 - 97 % 2 - 3.5 % 1 - 2 %
Thnh phn hemoglobin ca Thnh phn hemoglobin ca
ngui ln ngui ln
Chc nng Hemoglobin Chc nng Hemoglobin
- Vn chuyn oxy dn m
- Phn ng gia Hb & oxygen:
- $ gn oxy - khng phi phn ng oxy ha.
- Mt Hb c th gn 4 phn t O2.
- $ gn oxy di hi t hon 0.01 giy.
- Chu nh hung ca mt s cht: 2,3-DPG
(2,3-Diphosphoglycerate).
Hb Hb - - Tnh ch Tnh cht (1) t (1)
t hp thun ngh[ch vi oxy tao oxyh0mogIobin
Hb + O
2
HbO
2
thm tuoi
- Su kt hop v phn ly gia O
2
v Hb duoc xc djnh bi
phn p oxygen mi trung chung quanh Hb.
phi: phn p O
2
cao F 95-98% Hb kt hop oxy F theo
mu dn m.
m: phn p O
2
thp (40mmHg) phn ng xy ra theo
chiu nghjch, HbO
2
phn ly, nh O
2
cho m
Fchc nng ca Hb trong h hp.
ung cong phn ly ung cong phn ly
oxy oxy - -hemoglobin hemoglobin
- $ kt hp v phn ly gia O2 v Hb
duc xc dnh bi phn p oxy (pO
2
)
- C dang $igma
- Vic gn oxy vo hem th nht lm d dng
gn oxy vo hem th 2 (co ch hip dng-
cooperative)
- !50 (phn p O2 cn thit d 50% Hb bo
ha vi O2) 26.8mmHg
ung cong phn ly Hb ung cong phn ly Hb- -oxy oxy
(dung cong Barcroft) (dung cong Barcroft)
Phn p oxy
Tnh mach ng mach
ung cong phn ly Hb ung cong phn ly Hb - - oxy oxy
phn PO
2
thp (noi HbO
2
nh O
2
cho m): dung cong rt
dc 1 su thay di nh ca PO
2
s lm thay di su giao O
2
cho m mt cch dng k gip hng djnh PO
2
/m.
phn PO
2
cao (noi Hb ly O
2
t phi) dung cong t:
PO
2
: 60mmHg F bo ha Hb vi O
2
: 89%.
PO
2
500 th d bo ha HbO
2
cng chi 100%
F PO
2
mi trung, ph nang c th thay di nhiu nhung d bo
ha HbO
2
& luong oxy giao m khng bj giao dng lm.
PO
2
m n djnh.
Hb c tnh dm gip PO
2
/ m n djnh, dm bo su hng djnh
ni mi.
- V tr bnh thung ca dung cong
ty thuc vo:
Nng d 2,3-DPG
Nng d ion H
+
(pH) (Bohr effect)
CO
2
trong hng cu
Cu trc ca Hb.
ung cong phn ly Hb ung cong phn ly Hb- -oxy oxy
Oxy & deoxyhemoglobin Oxy & deoxyhemoglobin
- Lch phi (d nh oxy)
- 2,3-DPG cao
- H
+
cao
- CO
2
cao
- Hb$
- Lch tri (d gn oxy)
- 2,3-DPG thp
- HbF
ung cong phn ly Hb ung cong phn ly Hb- -oxy oxy
Hb Hb -- Tnh cht (2) Tnh cht (2)
t hp vI carbondioxyd (CO

tao carbonyI Hb (CO

Hb
CO
2
: Sn phm chuyn ha cc mFthi ra ngoi
Mu TM: 50-60ml CO
2
/100ml mu
Mu M: 50-53ml CO
2
/100ml mu.
PO
2
= 40mmHg, 37
o
C, chi 2,9ml CO
2
ha tan F phi vn chuyn bng nhiu cch.
a) Hb kt hop truc tip vi CO
2
: qua nhm NH
2
tu do ca Globin tao thnh carbonyl Hb(HbCO
2
)
R-NH
2
+ CO
2
R-NH-COOH
(dn xut carbamyl)
P/u xy ra F hoc ty thuc PCO
2
m : PCO
2
= 46mmHg : phn ng theo chiu thun
phi: PCO
2
= 36mmHg : phn ng theo chiu nghjch
b) Hb vn chuyn gin tip CO
2
:
CO
2
thm thu t cc m vo mu:
CO
2
+ H
2
O H
2
CO
3
H
+
+ HCO
3
-
CA ( carbonic anhydrase)
H
+
gn vo His (146) tn cng chui ca Hb tu do (va gii phng O
2
) tao Hb2H
+
F dn phi: H
+
duoc gi phng + HCO
3
-
H
2
CO
3
.
H
2
CO
3
H
2
O + CO
2

CA
Th ra
2CO
2
+ 2H
2
O
2H
2
CO
3
2HCO
3
-
+ 2H
+
4O
2
Phi
Hb.4O
2
Hb.2H
+
4O
2
2H
+
+ 2HCO
3
-
2H
2
CO
3
2CO
2
+ 2H
2
O
Sf ca
CT Krebs
M
SO
'N CHUYN CO
2
GN TP
CA Hb
CA
CA
(Carbonic
Anhydrase)
Hb Hb -- Tnh cht (3) Tnh cht (3)
t hp vi Oxyd carbon (CO tao carbon
monoxyd Hb (HbCO) rt bn vng.
CO c i luc vi Hb gp 210 ln so vi oxy
CO c th dy O
2
ra khi HbO
2
Hb + CO HbCO
HbO
2
+ CO HbCO + O
2
Trong trung hop ng dc CO:
Hb Hb Tnh cht (4) Tnh cht (4)
4.Oxy ha Hb tao M0th0mogIobin (M0tHb
cht oxy ha
Hb MetHb + e
-
(Fe
+2
) (Fe
+3
)
nitrit, clorat,
ferricyanua.
Hb Hb Tnh cht (5) Tnh cht (5)
Tnh cht 0nzym
Tnh cht ca mt peroxydase:
H
2
O
2
+ AH
2
2H
2
O + A
Hb
Tnh cht ca mt catalase:
H
2
O
2
2H
2
O + O
2
.
Hb
Kt lun Kt lun
W Gim hemoglobin trong hng cu do bt
ky bt thung no trong bt ky thnh phn
no ca Hb du dn dn tnh trang thiu
mu (anaemia)
W T d lm thiu oxy cung cp cho m.
%ng hp chui Globin %ng hp chui Globin
.|usler - .rorosore 1
.|usler - .rorosore 11
2s2 0oWer 1
22 Porl|ard Eroryor|.
2s2 0oWer ll
22 F Fela| <1
2o2 A2 1.5-3.5
Adu|l
22 A >95
l 2
s C o A
hemog|ob|n e|ectrophores|s:
Variants of sickle cell anemia
hemog|ob|n e|ectrophores|s:
/entification of abnormal hemoglobins