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III with WSXWS
IRS IRS-binding SOCS SOCS-binding
STAT
binding
STAT
binding
STAT
binding
SHP
STAT STAT-binding
binding
Jak 2
. Tyk 2
JAK/TYK -binding
Jak 1
SOCS SHP
STAT
Rho-GEF homology domain
. Sprouty
Sprouty domain
PTB
PTB domain
PH
4 Plekstrin homology
domain
Protein separation using SDS-PAGE
(Laemmli system)
1. Apply protein/dye samples into polyacrylamide gel wells
.
6
Embryonic stem cells
Sperm donor Egg donor
Egg fertilization
Culture fertilized cells for several cell divisions
Dissociate cell cluster for subculturing and
1
2
3
IRS
SOCS
STAT
binding
STAT
binding
FN Fibronectin III type III domain
FN Fibronectin
CCCC type III domain
III with cysteines
STAT
binding
FN Fibronectin
WS-WS type III domain
NO – mediated attenuation of adrenergic Gq signaling in VSMC
ANP
K+ BKCa
pGC
sGCCa
cGMP
PKG
PLC
q
GTP
q
GTP
PA
1,4,5-Ins
PKC
q
GDP
Ca Ca
Ca Ca Ca
Ca
Ca Ca Ca
Ca Ca Ca
Ca Ca Ca Ca
BLNK
SLP-76
SHC-1
APS
845
SH3
Vav-1
SH3
Src-homology 3 domain
SH2
Src-homology 2 domain
SAM
C-1
SAM domain
Cysteine-rich region 1
Calponin
Calponin homology domain
Rho-GEF
III
FN
WS-WS
III FN
III
FN
III
FN
III
LIF-R 1053 aa
FN
CCCC
III FN
WS-WS
III
Ig
FN
CCCC
III
FN
WS-WS
III FN
III
FN
III
FN
III
Tyk Jak
2
1
STAT
binding
Jak Tyk
1
2
Jak Jak
Jak
1
2
1
Jak
Tyk
Jak
OSM-R 952 aa
IL-12 662 aa
IL-6R 449 aa
IL-11R 400 aa
FN
CCCC
III
FN
WS-WS
III
FN
Ig Ig III
FN
CCCC
III
FN FN
CCCC
III III
FN FN FN
WS-WS WS-WS
III
III III
FN
WS-WS
III
Ig
FN
CCCC
further utilization
Potential use for
therapeutic cloning .
Use for In Vitro fertilization
Freeze in N2 for
long-term sto7rage
2. Run the electrophoresis until dye reaches the end of the gel
Stacking gel
Resolving gel
3. Remove the gel from the apparatus and stain for proteins
.
5
A Column Chromatography Toolkit
IL-10R2 325 aa
III
FN FN
FN FN
III III
III III
FN FN
FN FN
III III
III III
INFR1 489 aa
IL-22R1 574 aa
INFR2 332 aa
FN FN
FN
III III
III
FN FN
FN
III III
III
IL-13R1 427 aa
IL-4R 825 aa
Ig
FN
CCCC
III
FN
WS-WS
III
Ig
FN
CCCC
III
FN
WS-WS
III
IL-13R2 380 aa
c 349 aa
Ig
FN
CCCC
III
FN
WS-WS
III
Ig
FN
CCCC
III
FN
WS-WS
III
gp130 896 aa
Ig
FN
CCCC
IFN III FN III
cAMP cAMP
cAMP cAMP
cAMP
cAMP
cAMP
cAMP cAMP
cAMP
PKA
14-3-3 P
RGS
14-3-3
.
2
Modular Structure of Cytokine Receptors
IFNR1 530 aa
IL-10R1 557 aa
FN
III
IFNR2 515 aa
FN
binding
STAT
binding
Jak
Jak
3
3
STAT
binding
STAT
binding
Jak 3
STAT
binding
STAT
binding
SHP SHP-binding
3
Ig Ig - like
Adaptor proteins - selected (human)
1
SH3
SH2
217
SH3
1
276
SH3
SH2
1
SH2
SH3
304
SH3
1
SH3
SH2
330
SH3
1
SH3
SH3
SH3
377
SH2
1
Sprouty
456
SH2
1 SAM
533
SH2
1
PTB
584
SH2
1
632
PH
SH2
1 Calponin
Rho-GEF
PH
C-1
SH3
SH2
GRB2
SLAP
Crk
GADS (GRID)
Nck1
Ca
Ca Ca Ca
Ca
Ca
Ca CaM
Ca Ca
Ca Ca Ca
Ca
MLCK
Ca
Ca
Ca
.
Ca
CaM
Ca
P MLCK
P
myosin
myosin
P
myosin
1
RGS function and regulation
Ligand
S
GDP RGS
TNF
Ligand
S
GTP
RGS
P
RGS
S
GTP AC AC
IRS IRS-binding SOCS SOCS-binding
STAT
binding
STAT
binding
STAT
binding
SHP
STAT STAT-binding
binding
Jak 2
. Tyk 2
JAK/TYK -binding
Jak 1
SOCS SHP
STAT
Rho-GEF homology domain
. Sprouty
Sprouty domain
PTB
PTB domain
PH
4 Plekstrin homology
domain
Protein separation using SDS-PAGE
(Laemmli system)
1. Apply protein/dye samples into polyacrylamide gel wells
.
6
Embryonic stem cells
Sperm donor Egg donor
Egg fertilization
Culture fertilized cells for several cell divisions
Dissociate cell cluster for subculturing and
1
2
3
IRS
SOCS
STAT
binding
STAT
binding
FN Fibronectin III type III domain
FN Fibronectin
CCCC type III domain
III with cysteines
STAT
binding
FN Fibronectin
WS-WS type III domain
NO – mediated attenuation of adrenergic Gq signaling in VSMC
ANP
K+ BKCa
pGC
sGCCa
cGMP
PKG
PLC
q
GTP
q
GTP
PA
1,4,5-Ins
PKC
q
GDP
Ca Ca
Ca Ca Ca
Ca
Ca Ca Ca
Ca Ca Ca
Ca Ca Ca Ca
BLNK
SLP-76
SHC-1
APS
845
SH3
Vav-1
SH3
Src-homology 3 domain
SH2
Src-homology 2 domain
SAM
C-1
SAM domain
Cysteine-rich region 1
Calponin
Calponin homology domain
Rho-GEF
III
FN
WS-WS
III FN
III
FN
III
FN
III
LIF-R 1053 aa
FN
CCCC
III FN
WS-WS
III
Ig
FN
CCCC
III
FN
WS-WS
III FN
III
FN
III
FN
III
Tyk Jak
2
1
STAT
binding
Jak Tyk
1
2
Jak Jak
Jak
1
2
1
Jak
Tyk
Jak
OSM-R 952 aa
IL-12 662 aa
IL-6R 449 aa
IL-11R 400 aa
FN
CCCC
III
FN
WS-WS
III
FN
Ig Ig III
FN
CCCC
III
FN FN
CCCC
III III
FN FN FN
WS-WS WS-WS
III
III III
FN
WS-WS
III
Ig
FN
CCCC
further utilization
Potential use for
therapeutic cloning .
Use for In Vitro fertilization
Freeze in N2 for
long-term sto7rage
2. Run the electrophoresis until dye reaches the end of the gel
Stacking gel
Resolving gel
3. Remove the gel from the apparatus and stain for proteins
.
5
A Column Chromatography Toolkit
IL-10R2 325 aa
III
FN FN
FN FN
III III
III III
FN FN
FN FN
III III
III III
INFR1 489 aa
IL-22R1 574 aa
INFR2 332 aa
FN FN
FN
III III
III
FN FN
FN
III III
III
IL-13R1 427 aa
IL-4R 825 aa
Ig
FN
CCCC
III
FN
WS-WS
III
Ig
FN
CCCC
III
FN
WS-WS
III
IL-13R2 380 aa
c 349 aa
Ig
FN
CCCC
III
FN
WS-WS
III
Ig
FN
CCCC
III
FN
WS-WS
III
gp130 896 aa
Ig
FN
CCCC
IFN III FN III
cAMP cAMP
cAMP cAMP
cAMP
cAMP
cAMP
cAMP cAMP
cAMP
PKA
14-3-3 P
RGS
14-3-3
.
2
Modular Structure of Cytokine Receptors
IFNR1 530 aa
IL-10R1 557 aa
FN
III
IFNR2 515 aa
FN
binding
STAT
binding
Jak
Jak
3
3
STAT
binding
STAT
binding
Jak 3
STAT
binding
STAT
binding
SHP SHP-binding
3
Ig Ig - like
Adaptor proteins - selected (human)
1
SH3
SH2
217
SH3
1
276
SH3
SH2
1
SH2
SH3
304
SH3
1
SH3
SH2
330
SH3
1
SH3
SH3
SH3
377
SH2
1
Sprouty
456
SH2
1 SAM
533
SH2
1
PTB
584
SH2
1
632
PH
SH2
1 Calponin
Rho-GEF
PH
C-1
SH3
SH2
GRB2
SLAP
Crk
GADS (GRID)
Nck1
Ca
Ca Ca Ca
Ca
Ca
Ca CaM
Ca Ca
Ca Ca Ca
Ca
MLCK
Ca
Ca
Ca
.
Ca
CaM
Ca
P MLCK
P
myosin
myosin
P
myosin
1
RGS function and regulation
Ligand
S
GDP RGS
TNF
Ligand
S
GTP
RGS
P
RGS
S
GTP AC AC