Nucleotide Analogs - Jena Bioscience

Recombinant Proteins
138
Store: -80 °C
Selected references:
Wenzel-Seifert et al. (2002) Similarities and differences in the
coupling of human β 1 - and β 2 -adrenoreceptors to G sα splice
variants. Biochem. Pharmacol. 64:9.
Wenzel-Seifert, K. and Seifert, R. (2003) Properties of Arg 389 -
β 1 -adrenoceptor-G sα fusion proteins: Comparison with Gly 389 -β 1 -
adrenoceptor-G sα fusion proteins. Receptors Channels 9:315.
Small et al. (2003) Pharmacology and physiology of human
adrenergic receptor polymorphism. Annu. Rev. Pharmacol.
Toxicol. 42:381.
β 2 -Adrenergic receptor CAM -G sαL
human,
Recombinant, Sf9 insect cells
Cat. No. Amount Price (€)
PR-531 1 ml 300,--
Membrane suspension. Supplied in 75 mM Tris-HCl,
pH 7.4, 12.5 mM MgCl 2 , and 1 mM EDTA.
β 2 -Adrenergic Receptor CAM -G sαL is a fusion protein in
which the G sαL N-terminus is linked to the β 2 AR CAM
C-terminus via a hexahistidine (His6)-Tag.
The human β 2 -adrenoreceptor (β 2 AR) is activated
by the catecholamine epinephrine and couples to
the G-protein G s to mediate adenylate cyclase (AC)
activation. β 2 ARs are found in numerous tissues and
cell types including vascular and bronchial smooth
muscle cells, leukocytes and liver. β 2ARs mediate
smooth muscle relaxation, inhibition of leukocyte
function and activation of glycogenolysis.
β 2AR CAM is a constitutively active mutant of the β 2AR.
It differs from the wild-type receptor by four discrete
amino acid substitutions in the third intracellular loop
of the receptor (L266 → S266, K267 → R267, H269 →
K269, and L272 → A272).
G sαL is the long splice variant of the α-subunit of the
heterotrimeric G-protein G s. G s activates the effector
AC. G sαL differs from the short splice variant (G sαS) by a
15-amino acid insert between the ras-like domain and
the α-helical domain. G sαL (cat.# PR-501) possesses a
lower GDP-affi nity than G sαS (cat.# PR-505).
The β 2AR CAM-G sαL fusion protein ensures a defi ned 1:
1 stoichiometry of the receptor and the G sαL subunit
as well as high coupling effi ciency. Compared to
the β 2 AR-G sαL fusion protein (cat.# PR-532), the
β 2AR CAM-G sαL fusion protein possesses an increased
constitutive activity as assessed by the increased
effi cacies of partial agonists and inverse agonists in
the steady-state GTPase assay.
The fusion protein contains a N-terminal FLAG-Tag ®
for immunochemical detection.
Store: -80 °C
Selected references:
Samama et al. (1993) A mutation-induced activated state of the
α 2 -adrenergic receptor. Extending the ternary complex model.
J. Biol. Chem. 268:4625.
Seifert et al. (2001) Functional differences between full
and partial agonists: Evidence for ligand-specifi c receptor
conformations. J. Pharmacol. Exp. Ther. 297:1218.
Seifert et al. (1998) Different effects of G sα splice variants on
α 2 -adrenoreceptor-mediated signaling.
J. Biol. Chem. 273:5109.
Seifert et al. (1998) Reconstitution of α 2 -adrenoceptor-GTPbindingprotein
interaction in Sf9 cells: High coupling effi ciency
in α 2 -adrenoceptor-G sα fusion protein. Eur. J. Biochem. 255:
369.
Wenzel-Seifert et al. (2002) Similarities and differences in the
coupling of human α 1 - and α 2 -adrenoreceptors to G sα splice
variants. Biochem. Pharmacol. 64:9.
β 2 -Adrenergic receptor-G sαL
human, Recombinant, Sf9
insect cells
Cat. No. Amount Price (€)
PR-532 1 ml 300,--
Membrane suspension. Supplied in 75 mM Tris-HCl,
pH 7.4, 12.5 mM MgCl 2, and 1 mM EDTA.
β 2 -Adrenergic receptor-G sαL is a fusion protein in which
the G sαL N-terminus is linked to the β 2 -adrenoceptor
(β 2 AR) C-terminus via a hexahistidine (His6)-Tag.
The β 2 AR is activated by the catecholamine
epinephrine and couples to the G-protein Gs t o mediate
adenylate cyclase (AC) activation. β 2 ARs are found in
numerous tissues and cell types including vascular
and bronchial smooth muscle cells, leukocytes and
liver.
β 2 ARs mediate smooth muscle relaxation, inhibition of
leukocyte function and activation of glycogenolysis.
G sαL is the long splice variant of the α-subunit of the
heterotrimeric G-protein G s . G s activates the effector
AC. G sαL differs from the short splice variant (G sαS) by a
15-amino acid insert between the ras-like domain and
the α-helical domain. G sαL (cat.# PR-501) possesses a
lower GDP-affi nity than G sαS (cat.# PR-505).
The β 2 AR-G sαL fusion protein ensures a defi ned 1:
1 stoichiometry of the receptor and the G sαL subunit
as well as high coupling effi ciency. Compared to the
β 2 AR-G sαS fusion protein (cat.# PR-544), the β 2 AR-G sαL
fusion protein possesses an increased constitutive
activity as assessed by the increased effi cacies of
partial agonists and inverse agonists in the steadystate
GTPase assay.
The fusion protein contains a N-terminal FLAG-Tag ®
for immunochemical detection.
Protein concentration: 0.5 – 2.7 mg/ml
Receptor expression level: 4.8 – 15.5 pmol/mg
Store: -80 °C
Selected references:
Seifert et al. (1998) Different effects of G sα splice variants on α 2 -
adrenoreceptor-mediated signaling. J. Biol. Chem. 273:5109.
Seifert et al. (1998) Reconstitution of α 2 -adrenoceptor-GTPbindingprotein
interaction in Sf9 cells: High coupling effi ciency
in α 2 -adrenoceptor-G sα fusion protein. Eur. J. Biochem. 255:
369.
Wenzel-Seifert et al. (2002) Similarities and differences in the
coupling of human α 1 - and α 2 -adrenoreceptors to G sα splice
variants. Biochem. Pharmacol. 64:9.
β 2 -Adrenergic receptor-
G sαL-Leu 227
human, Recombinant, Sf9
insect cells
Cat. No. Amount Price (€)
PR-533 1 ml 300,--
Membrane suspension. Supplied in 75 mM Tris-HCl,
pH 7.4, 12.5 mM MgCl 2, and 1 mM EDTA.
β 2-Adrenergic receptor-G sαL-Leu 227 is a fusion protein
in which the G sαL -Leu 227 N-terminus is linked to the
β 2-adrenoceptor (β 2AR) C-terminus via a hexahistidine
(His6)-Tag.
The β 2 AR is activated by the catecholamine epinephrine
and couples to the G-protein G s to mediate adenylate
cyclase (AC) activation. β 2 ARs are found in numerous
tissues and cell types including vascular and bronchial
smooth muscle cells, leukocytes and liver.
β 2 ARs mediate smooth muscle relaxation, inhibition of
leukocyte function and activation of glycogenolysis.
G sαL is the long splice variant of the α-subunit of the
heterotrimeric G-protein G s . G s activates the effector
AC. G sαL differs from the short splice variant (G sαS ) by a
15-amino acid insert between the ras-like domain and
the α-helical domain. G sαL (cat.# PR-501) possesses a
lower GDP-affi nity than G sαS (cat.# PR-505).
The exchange of Gln 227 to Leu 227 (Q/L mutation) in the
catalytic site abolishes the intrinsic GTPase activity
and increases the GDP-affi nity of G sα , resulting in a
constitutively active G-protein.
The fusion protein contains a N-terminal FLAG-Tag ®
for immunochemical detection.
Protein concentration: 1.5 – 2.5 mg/ml
Receptor expression level: 7 pmol/mg
Store: -80 °C
http://www.jenabioscience.com
Selected references:
Seifert et al. (1998) Different effects of G sα splice variants on α 2 -
adrenoreceptor-mediated signaling. J. Biol. Chem. 273:5109.
Graziano, M. P. and Gilman, A. G. (1989) Synthesis in
Escherichia coli of GTPase-defi cient mutants of G sα . J. Biol.
Chem. 264:15475.
Seifert et al. (1998) Reconstitution of α 2 -adrenoceptor-GTPbindingprotein
interaction in Sf9 cells: High coupling effi ciency
in α 2 -adrenoceptor-G sα fusion protein. Eur. J. Biochem. 255:
369.
Gille et al. (2003) GDP affi nity and order state of the catalytic
site are critical for function of xanthine nucleotide-selective G αs
proteins. J. Biol. Chem. 278:7822.
β 2-Adrenergic receptor-
G sαL -Leu 227 -Asn 295
human, Recombinant, Sf9
insect cells
Cat. No. Amount Price (€)
PR-534 1 ml 300,--
Membrane suspension. Supplied in 75 mM Tris-HCl,
pH 7.4, 12.5 mM MgCl 2 , and 1 mM EDTA.
β 2 -Adrenergic receptor-G sαL -Leu 227 -Asn 295 is a fusion
protein in which the G sαL -Leu 227 -Asn 295 N-terminus is
linked to the β 2 -adrenoceptor (β 2 AR) C-terminus via a
hexahistidine (His6)-Tag.
The β 2 AR is activated by the catecholamine epinephrine
and couples to the G-protein G s to mediate adenylate
cyclase (AC) activation. β 2ARs are found in numerous
tissues and cell types including vascular and bronchial
smooth muscle cells, leukocytes and liver.
β 2 ARs mediate smooth muscle relaxation, inhibition of
leukocyte function and activation of glycogenolysis.
G sαL is the long splice variant of the α-subunit of the
heterotrimeric G-protein G s . G s activates the effector
AC. G sαL differs from the short splice variant (G sαS ) by a
15-amino acid insert between the ras-like domain and
the α-helical domain. G sαL (cat.# PR-501) possesses a
lower GDP-affi nity than G sαS (cat.# PR-505).
GTP-binding proteins possess a highly conserved
aspartate residue in the NKXD motif that is critical for
high-affi nity interaction with GTP. In small GTP-binding
proteins, the D/N-mutation switches base-specifi city
from guanine to xanthine. The exchange of Asp 295 to
Asn 295 leads to an inactive G sα -mutant. However, an
additional Q/L-mutation in the catalytic site (Gln 227 →
Leu 227 ) that abolishes GTPase activity and increases
GDP-affi nity rescues protein function and induces
specifi city for XTP (cat.# NU-602) and XppNHp
(cat.# NU-403) relative to GTP and GppNHp (cat.#
NU-401), respectively. In contrast, the mutant is not
specifi c for XTPγS (cat.# NU-404) relative to GTPγS
(cat.# NU-412), probably because of conformational
alterations in the catalytic site by the γ-thiophosphate.
The fusion protein contains a N-terminal FLAG-Tag ®
for immunochemical detection.
Protein concentration: 1.3 mg/ml
Receptor expression level: 3.7 pmol/mg
Store: -80 °C
Selected references:
Seifert et al. (1998) Different effects of G sα splice variants on α 2 -
adrenoreceptor-mediated signaling. J. Biol. Chem. 273:5109.
Graziano, M. P. and Gilman, A. G. (1989) Synthesis in
Escherichia coli of GTPase-defi cient mutants of G sα . J. Biol.
Chem. 64:15475.
Seifert et al. (1998) Reconstitution of α 2 -adrenoceptor-GTPbindingprotein
interaction in Sf9 cells: High coupling effi ciency
in α 2 -adrenoceptor-Gsá fusion protein. Eur. J. Biochem. 255:
369.
Gille et al. (2003) GDP affi nity and order state of the catalytic
site are critical for function of xanthine nucleotide-selective G αs
proteins. J.Biol. Chem. 278:7822.