Các tính chát ca bán cua enzym

-Làm täng vân tóc phán úmg mà không làm thay dői tién trinh cúa phán úmg
+ Xét phán úng: sucrose + 02 cO2 +H20

- Phán úng (thuòng) èdiëu kiên nhę nhàng: t, pH, áp suát aEnA a
Dãc hiêu (cao) óicO chat & sán phäm
-

Ben chat:ARN, protein

stain bar nhn can sel
noto
chat
I nay by sinhly'
·

Hat to tiehi, nhay cam is No enz e

Thrombin
Trypsin Lys drolysls site Hydrolyais site
or

Argi
Time
 Enzym lam tang van toc phan ung

Rate
Catalyzed enhancement
Enzyme half-life rate (kun s-1)
OMP decarboxylase 78,000,000 years 2.8 X 10-16 39 1.4 X 1017
Staphylococcal nuclease 130,000 years 1.7 X 10-13 95 5.6 X 1014
AMP nucleosidase 69,000 years 1.0 X 10-11 60 6.0 X 1012
Carboxypeptidase A 7.3 years 3.0 X 10-9 578 1.9 X 1011
Ketosteroid isomerase weeks 1.7 x 10-7 66,000 3.9 X 1011
Triose phosphate 1.9 days 4.3 X 10-6 4,300 1.0 X 10°
isomerase

Chorismate mutase 7.4 hours 2.6 X 10-5 50 1.9 X 10%

Carbonic anhydrase 5 seconds 1.3 X 1071 1 X 10° 7.7 X 10t

Abbreviations: OMP, orotidine monophosphate; AMP, adenosine monophosphate.

Source: After A. Radzicka and R. Wolfenden. Science 267:90-93, 1995.
Enzym läm gläm năng luçng tų do hoąt hoá

Su thay dói năng lu ng tyr do Gibbs AG (kJ/mol)

= G sán phäm – G ca chát
S1 t.
– AGo'; dièu kiên chuân (S) = 1 M & pH7,0
Transition state, Xt

AG (uncatalyzed) ACGx Gs
-

Ac (catalyzed)
Substrate

AG
(or the
reacion)

Product
ATP +H20= ADP + P

Reaction progress -
(G0'=
AC -30,5 kJ/mol (-7,3 kcal/mol) ↓ lamtn thoutho
Enzym thúc dáy phán úng nhanh chóng dąt cân bång (equillbrium)
Phán úng dat trang thál cán báng dông hoc khi không có enzym: S

4
Häng ső cân bâng cúa phán úmg: P k 10
10
K = 100
(S) kR
+Enzyme

No enzyme

Seconds

Time-
Trung tâm hoąt dông cua enzym (active site)
le chain
sic
– Noi enzym két hop vói co chát & chuyén thanh
sán phäm cúa phán úng I

enzym
Thuòng giőng khe nút/ ränh trên bè măt
khoáng không gian cho ca chát liên
0 mach chink
main chain

két vól enzym

-
&
- Turong tác giūra cd chät & trung tâm hoąt dông
= các liên két yêu

+Liên két hydro

-chix a bhiptgan nhau
+Liên két van der Wallls
=>
In that
you
+ Turang tác ky nu c Lysozym (pdb ID 6lyz) ase
hat.
+ Tuong tác tính diên
008 88 C

I
+Liên két công hoá trj thuân nghjch 35 52 62,63 101 108 129

hotatc tao thmuch nhan

-

Try tain
hot lainam xanhem?
often
teis cas
⑪
an

↳ Vi

⑪ Tai sar can Gaa chottam hot

inacantain 123aa cho team

mach

fais.hijccheit'
⑧ vatar hot linkli
gei?
Lièn két hydro giüa co chát & trung tâm hoąt dông
Eugenol oxidase Ribonuclease

R278
Y168
UracilR
q425 M282 (from
substrate)
1427 E378
15l
V436

V166 C
Threonine
side chain

Y471

H390
G392
Serine
O R472 side chain
⑧
Y91

Datatin 0 Thi
Trung tâm hoąt dông cua eugenol oxidase
cochai.

I
Mó hinh chia khoá - khoá Emil Fisher 1894

Substrate

Active site

ES complex

Hermann Emil Fischer

(1852–1919)
1902 Nobel Prize in chemistry Enz mld
 IkH

0 0

Hain hot
glucobinase
an

⑩
yen
to nir pot tink dac hier enzym
⑧
This
glucobinase
tien
this
glucose
 so'ladag L
try cathe" sa
 -

khi nhorn
con

Ox:sxcicho puncio try cathe"

CO2 toes the
trichonianty othe
 ATE.
↳sitgic
 ganchatvrenzim
gan'ranheinen ↓
Cofactor là lon kim loai Metalloenzymes
Carbonic anhydrase Simulated
lons Enzyme5
(CA) enzyme CA active site
Cu+ Cytochrome Oxidase
H,0
Fe or Fe Cytochrome oxidase, catalase,
peroxidase
K Pyruvate kinase

Mg"+ Hexokinase, glucose 6-phosphatase,

p ruvate kinase
Mn+ Arginase, ribonucleotide reductase
CA-Zn (H, )
Mo Dinitrogenase CO
N4 rease

Zn* Carbonic anhydrase, alcohol

H,0
dehydrogenase, carbox peptidases
A and B 10,
CA-Zn (HCo,)- CA-Zn O )
22
 Tyr 96
Phe 87

Val 247

Asp 297

Leu 244

Camphor (substrate)

Val 295

Heme
Ky thuât djnh luçng enzym, ví dų alcohol dehydrogenase (ADH)

10 NAD
(dang oxy ho )
a8
Ficohthat
NADA
O.66 NADH
VADA
0X (dąng khu)

02

00
io 2) * 360) 380

Buóe sóng (nm) EB

philiman
that itsthis
= toictiti
~Go
nach
be
bin sie
puoxiclare
Ky thuât djnh luçng enzym két hop
Glucose Oxidase Cluc0se
ATP Mg2
-D- lucU▇ Mutaro -D-GlucDs
0. Hexokinase
glucose oxidase
ADP, Mg2
Gluconic acld H,0,
Gilucose-6-phosphate

Peroxidase
NAD)
,0, chromogen 7 Color complex
( -dlunisldln ) H * Glucose-6-phosphate
(phenl mln ) A 520 nm dehydrogenase 1 Grexokinage
Eutei

NADPH+H
6-Phosphogluconolactone

VG 0 Vin= Utes sink mu NADPH

E
 S
s,Estonistas
[I] S 10mM
=

A E +

C P his
5mM
&
S
=

ba.[ES]
7
S 1mM
= > Vi
S

⑧ t
⑧ t
[E]:const ⑧
Trag thai or dink StudyState
r
-
dis.
=

O
=k,.S:k,(so-p)
~ tho thanh ES R,[E]. [S]
=

2
~
maici (k,+ke). [ES]
=

4 h, [E]. [S]:(4, +he) [ES]

([ES]:
W
E

[E]:[ED+ [ES] ⑧
-

[S]
kit. [S]
⑧

->
hy[E]+.[S] V.(S]
enzymat be han
v=
km njat
(n chat tai co
kn*[3]=
-
so ate to
kn [S]
ke
+

+
[S]
km =

Umax Rcat. [EJt

=

V limp
=

[S] ->
e

4
⑧ Yrghiakm-- kmaythapthis aily'slay cas.

·Factor run+ AbH, cHICHONADH t

~
As his enzyme cochet.
CHICHO NAD+HO+

CHCOOH NADH+H
+

aihi hi.
>
Glucose 0.04
=

mM- ADLH

Hexohineekm km this (by the-> agacious tot,

Fructose:1,mM A DLH

km Cao
(bth)

M
Cam
=> te km inKat

meX /ts/1mmenz Bla

S
Lién quan giüa Vo & (co chát), (enzym)

Substrate concentration S Substrate concantration S

 ④ Cothi

Mó hinh dóng hoc Michaelis-Menten & Steady-state kinetics (1913)

Vmax ISJ Vo = Vmax

E + S ES E+P

(S)
Vo
= VmS + K M
VVmax k- + ky
-

K =
k

( ( l
Vma% = keatEJT = k ET

KM Nöng dò c chat |S)

k.
(cat V.ma ET
5/s
Dò th| Lineweaver-Burk (double-reciprocal plot) 1934
|SJ
Vo = Vm S) + KM
hixt
hager
Hans Lineweaver
(1907-2009)
/V, Slope =KM/Vmu

Intercept =–1/KM

Intercept =1n

Substrate concentration S Dean Burk

(1904–1988) 34
Km &ý nghīia trong chuyén hoá
AV
Enzyme Substrate K(mu)
Hexokinase (brain) AP 0

D-Glucose 0.06
Pructose
V AVA cha
HCO, 26
tytrig ein
nat Carbonic anhydrase

Sie Chymotrypsin Glycyltyrosinylglycine 108

as
N-Benzoyltyrosinamide 25
AS AS
-Galactosidase D-Lactose 0I)

Threonine dehydratase L-Threonine D.0

S).

Alcohol dehydrogenase
CH CH,OH + NAD = CH,CHO + NADH+ H+

Aldehyde dehydrogenase
CH,CH + NAD+ + H,0 CH,c00+ NADH 2H
Kcat = Vmax/E: turnover number, catalytic constant

E + S ES E + P
Turnover number

Enzyme (per second)

V. k|
(E
Carbonic anhydrase 600,000

3-Ketosteroid isomerase 280,000

E+ S EP E+P Acetylcholinesterase
Penicillinase
25,000
2,000

(Lactate dehydrogenase 1,000

Chymotrypsin 100
Vma = k E
DNA polymerase l 15

Tryptophan synthetase 2

Vo (maz/S)
V.
= Km + (S) Vo
k ES Lys0zymea 0.5

m Km + S|
6
 Cofactor FAD dóng vai trò ch t vàn chuyên
e quan trong Mô hinh d ng h c Michaelis-Menten & Steady-state kinetics (1913)
vongisoalloxazin
H+ CH Vo=Vmax
E+S ESE+P
FMN
CH
HCOR
HCOH
FADH (FMNH)
(dangsemiquinon)
FADH2 (FMNH2)
(dang khir hoàn toàn)
S]
V-VS]+Ku
HCOH

FAD --
H
- - - - -Riboflavin =vitamin B2
FAD flavin adenin

FMN
dinucleotid

flavin mononucleotid y2Vmax

COO
=
K
FAD FADH H CO0
H-¢-H VmaxkalE}T k:[E}rT

F-H 00C H
k-VmEr
sUccinat

OH Co0 dehydrogenase
KM Nong co ch t [s]-
Succinat Fumarat

Kcat= Vma/[E]: turnover number,catalyticconstant Keat/Km: catalyticefficiency,specificity constant (Hàngs c hi u)

E +S ES E +P Khá näng
acid ester
thuý phân c a a-chymotrypsin vói các N-acy--amino

Turnover number kcalE l[S

max ka[E,]
Enzyme per second) VoKn + [S] Amino acid in ester Amino acid side
chain ka/KM (s-l M-1)
Carbonic anhydrase 600,000
Glycine -H 1.3 X 10-1
3-Ketosteroid isomerase 280,000
CH
E +SESK- BP E +P Acetylcholinesterase
Penicillinase
25,000

2,000 Khi [S]<< Kmv

Valine CH 2.0

Lactate dehydrogenase 1,000 CH2

Norvaline 3.6X 102
Chymotrypsin 100 -CH,CH,CH,
maxka[E,]
DNA polymeraseI 15 Norleucine
=CH,CH,CH,CH, 3.0x10
2
CH2
Iryptophan synthetase

VmaxS] E ]S Lysozyme 0.5

Vo KAm (ES
Kcat Phenylalanine 1.0X 105

VoKn+S
cat
Vo
Kn +[S]
A Guide
and (W H Freeman
(W.H. Freea and dnsm in rotein Science:
Company, 1999).Table
Enzyme
12
Enzym di lap the (allosteric) khong theo mo hinh Michaelis-Menten

Multiple subunits & multiple active sites

VI du: aspartat transcarbamoylase (ATCase)

Michaelis-Menten kinetics Allosteric enzyme kinetics

ve l o c i t y ve l o c i t y
Re a c t i o n Re a c t i o n
Substrate concentration [S] Substrate concentration [S]
Anh huóng cua tº dén hoąt dò enzym–Nguyên nhân?
bein chai ez laprotein

Nononzymalio
chemieal
o)

Tomparaturo
optlmum for
0 (nzzmo+a ml.
(00
0cilon
AclivO enzymo
80

60

Enzym0-catalyzed
20
oaction

I) 20 30 A0

Temperature (C)
Anh huóng cúa pH dén hoąt dò enzym–Nguyên nhân?
Urease (urê C02 +NH )

pH 6.7

Cilrato
day
Pepsin pH 6.3 pH 7.3
Acotate
Phosphato
(Argini

Acetate.

Phosphat

Cilrate -
hotenz
He spHah-
Úc ché enzym
- Chát úe ché: làm giám ván tőC xúc tác cúa
H.
enzym

Bán chät:

+Chát chuyén hoá cúa té bào binh thuóng

+ Chat ngoąi sinh, chāt doc

Mt ső loal úrc ché enzym cochai gen vs ez:11:

1KCH5
Dihyydrofolat

HN.
1/ Không thuân nghijch: không thé loai bó chát
úrc chē bâng thäm tách
2/ Thuân nghjch: có thé loai bó chát úc ché
báng thäm tách
- Canh tranh

Khöng canh tranh

Methotrexat
 Mombrano phosphollpld
Töng hop eicosanoids tů acid arachidonic
Phospholpa o A2
1Clucocorticoid

Gyt C0o

Epoxidos 450
) Ae Arachldonla acld
(Cz04,
co0

Gyclo-0rygenan
LTA Aspirin and other NSAID5

Q
c0OH C00

PGH
OH (
TXA,
Thromboxanos

PGE PGF PGA PG

prostncydin)

Prostnglandi
Úc ché bång cách lièn két công hoá tr_Covalent inhibitors (1/2)
GOX-1
GI

nym0
Asplrin Inaativatod
Catolytle Slto u

t posltion 629 co0

Acotylatlon
0-cGH c0X-2
0-GCH
witehud
catalyilc 0ctivity

With anpirin

3
arine
A A

Channel of cid
U

Catelle-LLawSon (2001)
Plotolo1r N EngJMed;
345:1809.-1817
Úc ché bång cách lién két công hoá tr|_Covalent inhibitors (2/2)
A. Normal renctlon of acetylchollnesteraso

Acetylehollne Cholline Acetato

HC–C-o-CH,-CH,- N(CH) H –CH, -CH,-N(GHi) H,C– (o=0

(0

O 0 C–CH H,0 O

Enz Sor. Enz Sor Enz-Sör

B, Renctlon with organophospharus Inhibitors

CH CH.
e–01

O CH, (C ,
CH, CH, Inactive
Enz -Ser H-C-0- - 0 -C H
U L1

Diisopropylphosphofluoridat CH, CH

(DIPF) Enz Se
2/ Chát tuong dóng trang thál trung glan–Substrate analogs, sulclde inhibitors
Vnhbl
voup ihizolidin
ting

Peptidoglycan
(CH

CH

Co0
Reactive peptide
bond in
actam ting

↳ bictibhas there'
Lislln

OH CO0
Ser

Glycopepo Penicillloyl-enzyme complex

peptidase (enrymatically inacive)
27 Chát tuong dóng trąng thái trung glan–Substrate analogs, sulclde Inhibitors
A aMP * (Eunino
(OH

XanlliU
H0.
Ap yp xantliine
Inhibited
)4 i
bynllopurinol

Mo = S Md
SH

Uta
H0
H-0 i0 H,0 H20

Hypo annlu Xanihine

Xanthino-ens
complox

OH OH

anthine
oridaso
Alloxanlhino LA Inactivo
*LL
L h

Allopurnb Alloxanthine
(ox ypurinol) (:
3/ Kim loal näng gån (không däc hiêu) vào nhóm chúe enzym nhu–SH/eystein

Ferrochelatase
Hemoglobin Protoporphyrin IX + lron
&

Accumulates In RBC8
Fluoresces
ALA synthase
5-ALLA Succinyl CoA
Elevated in urine + Glycine
anndd plasma
Coproporphyrinogen
AA Oxidas6

dehydratase

Porphobilinogen -—Uroporphyrinogene C o p ro p o r p hy r i n o g e n
Elevated in urine
Cac co che uic ché enzym thuan nghich
chic chigan
hitchain
Uncompetitive
(A) Substrate (C) Substrate inhibitor

vcanktank
is

Enzyme Enzyme

(B) Competitive
inhibitor~
(D) Substrate.
Noncompetitive
inhibitor
so cheapthe
Enzyme
Enzyme

50
Üc ché canh tranh thuân nghjch (1/2)
C00
PAD FADH C00
H––H Substrate.
GH,
H-C–H
C Succinat (O0C
co0 dehydrogenase
(0
Succinat Fumarat
Malonat
Enzye

Competitive
inhibitor
S E+ P

K = (EJJ/EI Enzyme
Üc ché canh tranh thuân nghjch (2/2)

100
No inhibitor
Competitive 80
inhibitor = Ki

60
1NVo (= 10 K
No inhibitor 40
present I= 5 Ki

20

1/S
Substratej
Üc ché thuân nghjch không canh tranh uncompetitive

E4 Substrate-
Uncompenv
inhibitor
ES + |E + P

100
No inhibitor
ESI
60

+Uncompetitive
inhibitor

(= 10 K 5 K
20
No inhibitor
present

(Substrate)

K for uninhibited enzyme

Kh for = Kj

1S
Úc ché thuân nghjch không canh tranh (pure) noncompetitive
Substrate.
E | ES E+ P Noncompetitive
inhibitor.

E ESI

100
No inhibitor
No co l
80
inhibitor

60
1Vo = K

No
present
20 |- 10 K I5 K

(Substrate)
/S
K
Dièu hoà hoąt dông enzym bång co ché feedback

Si

== D

Gene transcription
Feedback inhibilion

Enzyme 3 Enzyme 4

Enzyme 2
Enzymo
Enzyme 6
Enzyme 6 *
roduct F
inhibilion 5
Dièu hoà hoąt dông ATCase bång ca ché feedback

(OP0,3- HN (C00 co0

Carbamoyl Aspartate N-Carbamoylaspartate

Hd (OH
phosphato
Cyildine triphosphate (C P)

ATCase = aspartat transcarbamoylase

Aspartate
Aa, carbamoylaspartate – UMP - UDP UTP CP
carbamoyl phosphate o
ATP
S6
Dièu hoă dij l p thé åm tinh & duong tinh (allosteric modulators)
Negatlvo ullouterie contral

negalive substrato
rogulator

Mo raclio CCsllt
L iid

The aclive site no tonger

fits the substrate.

i 0 I ULE0
*R
positive
*1

navailablo
L A 5l

5 i1 9 DsE
Dièu hoà di ąp thé aspartat transcarbamoylase (ATCase)
C00 (0
0

H,N.C—H HN CH

H,N CH, (CHC00

co01
aspartat
transcarbamoylasSe
carbamoyl phosphat aspartat N-carbamoy| aspartat

2 mMAI

0.4 mMCP

0 0
5
(Aspartate), mM Asppartate), mM
(CCP, mMM 58
Thay dôi các liên két công hoá trj thuân nghjch
Su phosphoryl hoá & kht phosphoryl hoá

, 2ATP
Phosphorylase
kinase 2ADP
Protein with serine side chain

OH
HO– P–0–ADP
Proteln
phosphatase
Protein ATP
H,0 * kinas0 Phosphorylase b Phosphorylase a

dang ít hoat dông dang hoąt dóng hon

- -0-- ADP
2P
Phosphoproteln
2H 0
phosphatase 1
Phosphorylated proteln
 I
-Hemono oxygenase 145t ~ethand
Xenobiogics (theactio x IkaI No sink:bile, steroids e
CYRG E

Xenobiotics
-
ngaeisinh
OXI
4
950
PHAI -

rhi R-H NADPH

+

H+
+

02
+ >
R-OH NADI
+
+
H2 0
>family (40%)

- man
thingfan ↓
-

cho e
<subfamily (>559)
↳hai e

Funchonalization
/

Gan
/NST, gene cluster
R -8H SO, Ihan be Riot, da, mame, pho nim
-

Glucomonid +than, Sertolle, han that

HAI
I lienhop Glutathion
Xenobiotics
Trogth
ER smooth
Glycin
V
Acetyl hoa'.
Majschai, may
tothe cam
lig Inhibitor
Chal it doe & tanUlO Indocer
⑰
V ⑦

7 CYIs" L

Ethanot: RM:0,03-0,04 mM gicii ctoi L -Actor tink.

Ethano Paracetamol
[ thismait
I

⑰ ~

104
ADI
⑦ NAD+
TCA ATB...
CHICHO
-

Misin
>
+

7 + NADH Ht
+ -

heattinh
(big
~
gan) gan -AcetylCoA CYPGE1.
A ·F. A acid bei
ALDH
CHICH8 +NAD+H+ X
CHICOOH +NADH +
Ht

<YM
aldehyd
>
L
man
-> 10.202 km< > mo/coquar ->Acetyl CoA NAIQI

CYLE1. (Microsomal ethanol enz-MECs) hoaiteigan

②
oxidizing
CHICHOH NADIH HT
+
+

02
+ >
CHCHO + NADIIHO GSH
W

Rm 11mM
=

gicictor
Hê mono-oxygenase cytochrom P450 trong chuyén hoá pha l
Function

Substrate functionalization
Oxidation or reduclion, Including aliphalic or NADPH NAD r, H

aromalic hydraxylation, opaxidalion, N, 0,

or S-deallylalion, Nhydroxylation, FAD
sulloxidation, desulluralion, oxidative RH0 ROH, H,0
dehalogenalion
FMIN
CHCH,O
Fo-hemo
Ethanol
NADPH
+H + 0

ethanokaxidizing NADP+
System (MEOs) + 2 0 Cytochrome Cytochrome
p450 reductase P450
0
C
H

Acetaldehyde
 SoI can
cytochrom P450 & b5 tai luoi n i sinh chât Nuoc buoi làm glàmchuyén hoá thuóc qua CYP3A4 th

Class IP4S0

REDUCTASE Fe,S P450

NADPH
FADFADH Fe Fe O+RHH,O+ROH
Hydroxylation
ClassI P450 Bergamottin

NADP)H
P450 REDUCTASE P450
Hydroxylation
acce OH

FAD PMN FMNH2 OgtRH HO+ROH

EAdalat
LA 60 CYP3A4

GF--4

C
Oytochrome b5 (afurocoumarin
CoA dimer)
b5 REDUCTASE Op+Oleoyl
NADH b5
FADFADH Stearoyl CoA+ H,O
Stearoyl COA desaturase

AUC T

-pharigicaO) pay at
P450 REDUCTASE P450
to
Atorvastatin

FADFMNFMNH O-RHHO+ROH
Hydroxylation Nifedipine Pharmacodynamics
Toxicity(
21 HMG-COA reductase Calcium channel blocker 22

Phenobarbital càm úng CYP2B làm t ng chuyên hoá thu c Chuyên hoá gây c cùa acetaminophen qua CYP2E1

-
Phenobarbtal
HN--CH N-C-CH
Kidney,
urine
UDPGA PAPS Kidney,
Sulfo transferase urine
UDP-glucurony
Increased transterase
oxidafive
expression of Glucuronate OH SOA
bioactvation activeCYP2B6
CYP Acetaminophen

EtOH CYP2E1
CAR
Cocain
Phenobarbital Increased
N-acetyi
transcription
of CYP286 mRNA cysteine

Plasma
HN-C-CH3 --cH HN--CHa
membrane GSH
Cell proteins
HN SG
Glutathione
S-protein
Stransferase
DNA OH OH
Hepatotoxicity
PBREM CYP2B6 Mercaptouric NAPQI
RXR acid
Nuclous -acetyl-p
benzoquinoneimine)
23 (toxic intermediate) 24
Kidney, urine
 Myc tiêu

Enzym& xúc tácsinh hoc Sau khi hoc xong chuong nåy sV có khá
n ng

1. & các thành phàn càn thift cho ho t d ng enzym

Trinh bày day dú bán chát protein cúa enzym

2. Giái thich d úng ca ch hoat d ng cia enzym thòng qua n ng lugng hogt hoá

3. Môtá ay các c i m cia trung tåm hoat ng cúa enzym

4. Phán tich úng các y u t ánh hu ng dén hoat ng enzym

Glucose Acid gluconic
s. Viet giái thích & tinh toán day dú các thòng s6 d ng hoc cia enzym theo mô hinh Michaelis-
+02 +H202
Menten

6. Giái thich düng khái ni m & cho vi du vë enzym di lêp thé

7. Phân tich dúng co ché, ánh huóng lên các thông s d ng hoc và de xušt úng dung tir các ca ché dc

ch enzym
Nguyen Quóc Thái,Pharm, PhD
Khoa Duoc- DHY Dugc Tp HCM 8. Môtá dây ú và phân tich cu th vi du vë các co ché kiém soát hogt ng cúa enzym
Emait nathai@ump.eduvn
9. Môtà phân vai trò c a h cytochrom P450 trong chuy n hoá thu c
Tháng 02/2023
loai, co ch ,

Phân loai enzym theo kiêu phàn úng xúc tác (1/3)
Trung tâm ho t ng c a enzym (active site)
1/ Oxidoreductase: xúc tác phán úng oxy hoá khú. Gôm 4 nhóm chinh:
Nai enzym két hop v í ca chát & chuy n thành

sán phám c a phán úng

-Oxidase, v: glucose oxidase, cytochrom c oxidase
- Thuong giong khe nút/ r nh trên bë m t
- Dehydrogenase, vd: alcohol dehydrogenase (ADH)
ADH
enzym khoáng không gian cho
coch t liên
- Peroxidase, catalase, glutathion peroxidase
vdt CHCHOH +NAD CHCHO +NADH +H*
ket voi enzym -Oxygenase göm: Ethanol Acetaldehyde

+Dioxygenase, vd: homogentisat dioxygenase

-Tuang tác gi a co chat & trung tâm hoat ng P450
+Monooxygenase, vd: cytochrom
các liênk t nào?
2/ Transferase: chuyén các nhóm chúc nhu amin, glycosyl, methyl, phosphory. Vi dur hexokinase

CH,OH Mg ATP CHOPOS

Lysozym (pdb
ID 6lyz)
OH ADP
NL 35
IL
52 62,63 101 108 29
H OH
Hexokinase

Gucose 6-phosphate 4
B-D-Glucose
 Phan loai enzym
3/ Hydrolase: phán

4/Lyase: phán úng

cát
theo kiéu

úmg thuy phán

liën

lien két oi. Vi du: pyruvat decarboxylase

C=O
liên

kEt C-C,-0,

CH3
Pyruvate
phànúng xúc tác (2/3)
kët

M
+H0

CN,
c-c,c-0,C-N,các

các

CO

Pyruvat

decarboxylase
lk
\
c ng hoá

CHs
Acetaldehyde
tri
lk công hoá

0 + HsN
tri

khác bâng cách

khác. Vi du:

loai
trypsin

nguyên t & tao

Phân logi enzym theo
5/ lsomerase:

6/ Ligase
chuy n
Vi du: malat isomerase, triosephosphat

-OH

CH,OPO,2

Dihydroxyacetone
phosphate

=synthetase: xúc tác phán

Succinyl
kiéu

Triosephosphat
isomerase

COA-

CoA
ph núng xúc tác (3/3)
nhóm chúc n i phân tù/ chuyén dong phán.

H--OH
isomerase

CH2OPO2
Glyceraldehyde
3-phosphate
(TIM)

úng tong hop có sy tham gia cúa ATP.

CH
COO
*
Succinyl

synthetase

+ ADP
CoA
CO0
CH2

CH2

coo
+ COA + ATP
Succinat
3 TIM barrel

Lactate Pyruvate
Cofactor+apoenzym = holoenzym/Coenzyme vs. prosthetic group Cofactor NAD(P)* ông vai trò
Dissociates COO
ch t v n chuyen e quan trong as H 0-G-H C0 +H
CH
CofactorsS "helper molecules
Alcohol

dehydrogena -NH2 -C-NH2

CHCHOH CHC-H
dân xu t cúa các vitamin)
Metal ions Coenzymes(thuong là
NAD NADH+H* Nicotinamide

NAD NADH
NAD H (dang oxy hoá) (dang kh)
öng co chat Cosubstrates Prosthetic groups Nhóm ngoai
********************|

NADP Bound AMP provides

Dnbound
dinding

CH that indu
conormadonal changes
the enZYme
O--
NAD

N=Hs H
OR
R«H
NADP
Niacinacid nicounic

itamin B

7 Nicotinamid adenin dinucleotid (phosphat)

 M t só co ché üc chê enzym Các co ché úc ché enzym thu n nghich
1/ Khongthuân nghich: không thé loai bó chät ütc ché bâng th m tách, vi du:
-Uc ché bång cách liëen két hoá tri-Covalent Uncompetitive
cong inhibitors
(A) Substrate () Substrate inhibitor

(COX)
Aspirin-cyclooxygenase

Disopropylphosphoftuoridat (DIPF)
-acetylcholinesterase
-Chat tuomg ong trang thái trung gian-Substrate analogs, suicide inhibitors
Enzyme Enzyme
Penicillin-gycopeptid transpeptidase
+Allopurinol xanthin oxidase (B) Competitive
-Kim loai n ng gån (không ëc hi nhóm chúc enzym nhu -SH/cystein inhibitor
u) vào (ví du ng c chl)
(D) Substrate
Noncompetitive
inhibitor.
2/ Thuannghich: có thé loai bá cht úcché bàng th m tách
-Canh tranh Enzyme
-Khong canh Enzyme
tranh

13
4

Uc ché canh tranh thu n nghich (1/2) Uc ché canh tranh thu n nghjch (2/2)

H
çoo
¢-
FAD FADH2 H C00 100
No inhibitor
CH2
H
Substrate +Competitive
H inhibitor
80
=K;
SUccinat
OOC
coo dehydrogenase 60
Succinat Fumarat 1/Vo
Malonat
yme No inhibitor
=10 K

Competitive
present
0=5 K;
ooooooocoooo 20
ES E+P inhibitor

1/[S]
Substrate]
Ki EJ11/(ED) K El
Enzyme
- Ibuprofen-
cyclooxygenase (COX)
- Statins
15 hydroxymethylglutaryl-CoA (HMG-CoA) reductase 16
Uc chéthu n nghich không canh tranh Uc ché thuan nghich không canh tranh
uncompetitive (pure) noncompetitive

Cyanid
E+ Uncompetitive
Substrate Inhibitor (D) Substrate cytochrom oxidase
ES+I E+P E+I ES E+P Noncompetitive
inhibitor -Arsenatglyceraldehyde

ESI Enzyme
No inhibitor
ESIX Enzyme
phosphate dehydrogenase

BO

Uncompetitive 100
inhibitor [0=K No inhibitor
EPSP synthase
+Noncompetitive
inhibitor
(-5K
oooocoes
No inhibitoor
present

Glyphosate
OH
0=10 Ki

Substrate
1/%|
No inhibitor
-K
present
KM for uninhibited enzyme 0-10K [=5K
o0oo0oooooooooo-
KPP for [I=K
Substrate
/[SI 17 1/[S]
18

Dièu hoà d th âm tính & duong tính (allostericmodulators)

l p
Hêmono-oxygenase cytochrom P450 trong chuyén hoá phal

Negative alosteric control Function

D
Substrate functionalization
negative
regulator substrate
Oxidation or reduction, including aliphatic or
NADPH NADP, H
aromatic hydroxylation, epoxidation, N, O,
No reactionoccurs. or S-deallylation, NAhydroxylation
active site
sulfoxidation, desulfuration, oxidalive
AD
enzyme
dehalogenation
AH Q2 ROH, H2O
The activesiteno longer
the substrate.
FMN
CHCHOH
Ethanol Fe-heme

Positive allosteric control

sUDStrate
NADPH
+H+O2 UDIDN
posilive Microsomal
reguialor
ethanol-oxidizing
NADP+
system (MEos) +2H20 Cytochrome Cytochrome
P450 reductase P450

cH
unavailable
acive
Enyne enzyme-substrate complex 19
Acetaldehyde