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5-FdUTP
Pharmacological modulation of cAMP
Forskolin
Ligand
Ligand
Ligand
Ligand


S
GTP ADP
S
GTP
i
AC AC GTP
i
GTP ADP




S
GDP NAD


i
GDP
ATP
cAMP
NAD
Cholera toxin
SAM
C-1
SAM domain Cysteine-rich region 1
Calponin Sprouty
Calponin homology domain Sprouty domain
Rho-GEF domain PTB
Rho-GEF homology PTB domain Plekstrin homology domain
cAMP
cAMP cAMP cAMP
cAMP cAMP
TNF
P
PKA
RGS
14-3-3
P 14-3-3
RGS
Modular Structure of Cytokine Receptors
OSM -R 952 aa gp130 896 aa IL-12 662 aa IFNR1 530 aa IL-10R1 557 aa IFNR2 515 aa INFR1 489 aa INFR2 332 aa IL-22R1 574 aa IL-4R 825 aa IL-13R1 427 aa c 349 aa IL-13R2 380 aa
pBluescript II
LacI
Ori
SP6 NsiI-HindIII-XbaI-EcoRI-SalI-SacI T7
T7
SfiI-SacI-EcoRI-SalI-Xho-BamHI-ApaI-XbaI-NotI-SphI-NsiI-HindIII
SP6
Βιβλιοθήκη Baidu
M13 (-)
A APCR
T 7
M13 (+)
Orotate phosphoribosyl transferase ATP Uridine kinase ADP
PRPP
Pharmacologically active compound 5-Fluorouridine 5’-nucleotidase 5-FUMP 5-FdUMP
5-FUTP
RNA DNA
eEF2
P
P
eEF2
eIF2
Ribosomal proteins
PABP eIF4G eIF4A eIF4E
40S
40S 30S
40S 30S
Mechanism of 5-fluorouracil (5-FU)
Thymidine phosphorylase
Uridine phosphorylase
Ig
FN
CCCC
III FN III FN III
Ig
FN
CCCC
Ig
FN
CCCC
III FN III FN III
III
III III
III III
III III
III III
III III
III
FN III
FN
WS-WS
FN
WS-WS
FN
WS-WS
FN
WS-WS
FN
WS-WS
FN
WS-WS
III
Thymidylate synthase
DNA synthesis
TTP
TMP
deoxyUMP
PH
Protein separation using SDS-PAGE
(Laemmli system)
1. Apply protein/dye samples into polyacrylamide gel wells
2. Run the electrophoresis until dye reaches the end of the gel
Cytidine deaminase
5-Fluoro-5-deoxy cytidine 5-Fluoro-5-deoxyuridine Thymidine phosphorylase
Capecitabine
Xeloda (prodrug)
Uridine phosphorylase
5-FU ATP 5-Fluorodeoxyuridine Thymidine kinase ADP 5’-nucleotidase
LIF-R 1053 aa
FN
CCCC
III III
FN
WS-WS
FN
WS-WS
III
Ig
FN
CCCC
Ig
FN
CCCC
Ig
FN
CCCC
FN
CCCC
FN III FN III FN III FN III FN III FN FN III FN FN III FN FN III FN FN III FN FN III FN
IL-10R2 325 aa
III III
FN
WS-WS
IL-6R 449 aa
IL-11R 400 aa
III III
III III
III III
FN
WS-WS
FN
WS-WS
FN
WS-WS
FN
FN
FN III FN
FN III FN III FN III
Ig
FN
CCCC
Ig
FN
CCCC
Ig
FN
CCCC
PDE
AMP
Pertussis toxin
Rolipram
Mechanism of Penicillin
(inhibition of crosslinking peptidoglycan)
Penicillin
Tetrapeptide
Glycopeptide transpeptidase
Pentaglycine
NO – mediated attenuation of adrenergic Gq signaling in VSMC
ANP
K+ BKCa
PLC
pGC
q
GTP


q
GTP

q
GDP
PKG
cGMP
PA
sGC Ca
1,4,5-Ins
PKC
Ca
Ca
Ca
Ca Ca Ca Ca Ca Ca Ca Ca Ca Ca Ca Ca Ca Ca Ca Ca Ca Ca Ca
Ca
CaM
Ca Ca
P MLCK
CaM
Ca
Ca
Ca Ca Ca
Ca
Ca
Ca Ca
MLCK
P
myosin myosin myosin
P
RGS function and regulation
Ligand Ligand


S
GTP
S
GTP
AC AC
cAMP cAMP cAMP cAMP

S
GDP RGS RGS
III
III
III
III
III
III
Tyk 2 STAT
binding
Jak 1
Jak 1
Tyk 2
Jak 1
Jak 2
Jak 1
Jak 1 IRS SOCS
Tyk 2
Jak 3
Jak 1 SOCS SHP STAT
binding
Jak 3
Jak 3 STAT
binding
Jak 3
STAT
binding
STAT
binding
STAT STAT
binding binding
STAT
binding
STAT
binding
STAT
binding
STAT
binding
STAT
binding
STAT
binding
FN Fibronectin III type III domain FN Fibronectin
456
SH3
1
SH2 SH2
533
BLNK
SH2
584
1 SAM 1
SLP-76
SH2
632
PTB
1 PH 1 Calponin Rho-GEF PH
SHC-1 APS
845
SH2
C-1
SH3
SH2
SH3
Vav-1
SH3 SH2
Src-homology 3 domain Src-homology 2 domain
5-FU
Carac, Efudex
5-Fluorodeoxyuridine Orotate phosphoribosyl transferase Uridine kinase Thymidine kinase
5-Fluorouridine
5-FUMP
5-FdUMP
DHF
N5,N10-methyleneTHF
Use for In Vitro fertilization
Freeze in N2 for long-term storage
Common Cloning Vectors
LacZ MCS
Amp Ori
r
f1 Ori LacZ Amp Ori
r
pBR322
4361bp Amp Tet r
r
MCS
pUC18
SOCS SOCS-binding
Ig
Ig - like
Adaptor proteins - selected (human)
1 217
SH3
1
SH2 SH2
SH3
276
GRB2 SLAP
304
SH3
1
SH2
1
SH3 SH2 SH3 Sprouty SH3
SH3
330
Crk
SH3
377
SH3
1
GADS (GRID) Nck1
CCCC
FN
WS-WS
III
IRS
Fibronectin type III domain with WSXWS IRS-binding
binding
SHP Jak 2 STAT STAT-binding Tyk 2 SHP SHP-binding
JAK/TYK -binding
III
type III domain with cysteines
Stacking gel
Resolving gel
3. Remove the gel from the apparatus and stain for proteins
A Column Chromatography Toolkit
Embryonic stem cells
Sperm donor
Egg fertilization
Culture fertilized cells for several cell divisions
Egg donor
Dissociate cell cluster for subculturing and further utilization
Potential use for therapeutic cloning
Inhibition of translation by Rapamycin (Sirolimus)
Growth Factor Receptors
Rapamycin
IP3K
PDK PKB
FKBP-12
mTOR
P
P
P
P P
4E-BP1
eEF2 kinase
P P P P P
CaM
P
S6K
P
4E-BP1 eIF4E
LacZ f1 Ori
LacZ f1 Ori
LacZ f1 Ori
pGEM 9Zf
2912bp Amp
r
pGEM 11Zf
Ori 3221bp Amp
r
pCR2.1
3929bp
Ori
Ori
r
Kan
Amp
Metabolism of capecitabine
Thymidine phosphorylase Carboxylesterase