RGSZ1, a G(z)-selective rgs protein in brain: Structure, membrane association, regulation by Gα(z) phosphorylation, and relationship to a G(z) gtpase-activating protein subfamily

Jun Wang, Axel Ducret, Yaping Tu, Tohru Kozasa, Ruedi Aebersold, Elliott M. Ross

Research output: Contribution to journalArticle

127 Citations (Scopus)

Abstract

We cloned the cDNA for human RGSZ1, the major G(z)-selective GTPase- activating protein (GAP) in brain (Wang, J., Tu, Y., Woodson, J., Song, X., and Ross, E. M. (1997) J. Biol. Chem. 272, 5732-5740) and a member of the RGS family of G protein GAPs. Its sequence is 83% identical to RET-RGS1 (except its N-terminal extension) and 56% identical to GAIP. Purified, recombinant RGSZ1, RET-RGS1, and GAIP each accelerated the hydrolysis of Gα(z)-GTP over 400-fold with K(m) values of ~2 nM. RGSZ1 was 100-fold selective for Gα(z) over Gα1, unusually specific among RGS proteins. Other enzymological properties of RGSZ1, brain G(z) GAP, and RET-RGS1 were identical; GAIP differed only in Mg2+ dependence and in its slightly lower selectivity for Gα(z). RGSZ1, RET-RGS1, and GAIP thus define a subfamily of G(z) GAPs within the RGS proteins. RGSZ1 has no obvious membrane-spanning region but is tightly membrane-bound in brain. Its regulatory activity in membranes depends on stable bilayer association. When co-reconstituted into phospholipid vesicles with G(z) and m2 muscarinic receptors, RGSZ1 increased agonist- stimulated GTPase >15-fold with EC50 <12 nM, but RGSZ1 added to the vesicle suspension was <0.1% as active. RGSZ1, RET-RGS1, and GAIP share a cysteine string sequence, perhaps targeting them to secretory vesicles and allowing them to participate in the proposed control of secretion by G(z). Phosphorylation of Gα(z) by protein kinase C inhibited the GAP activity of RGSZ1 and other RGS proteins, providing a mechanism for potentiation of G(z) signaling by protein kinase C.

Original languageEnglish (US)
Pages (from-to)26014-26025
Number of pages12
JournalJournal of Biological Chemistry
Volume273
Issue number40
DOIs
StatePublished - Oct 2 1998

Fingerprint

RGS Proteins
GTPase-Activating Proteins
Membrane structures
Phosphorylation
Brain
Association reactions
Membranes
Protein Kinase C
Muscarinic M2 Receptors
Proteins
GTP Phosphohydrolases
Secretory Vesicles
Guanosine Triphosphate
GTP-Binding Proteins
Cysteine
Hydrolysis
Phospholipids
Suspensions
Complementary DNA

ASJC Scopus subject areas

  • Biochemistry

Cite this

RGSZ1, a G(z)-selective rgs protein in brain : Structure, membrane association, regulation by Gα(z) phosphorylation, and relationship to a G(z) gtpase-activating protein subfamily. / Wang, Jun; Ducret, Axel; Tu, Yaping; Kozasa, Tohru; Aebersold, Ruedi; Ross, Elliott M.

In: Journal of Biological Chemistry, Vol. 273, No. 40, 02.10.1998, p. 26014-26025.

Research output: Contribution to journalArticle

@article{bbad9bf526bc4c06a9f9e41a50920031,
title = "RGSZ1, a G(z)-selective rgs protein in brain: Structure, membrane association, regulation by Gα(z) phosphorylation, and relationship to a G(z) gtpase-activating protein subfamily",
abstract = "We cloned the cDNA for human RGSZ1, the major G(z)-selective GTPase- activating protein (GAP) in brain (Wang, J., Tu, Y., Woodson, J., Song, X., and Ross, E. M. (1997) J. Biol. Chem. 272, 5732-5740) and a member of the RGS family of G protein GAPs. Its sequence is 83{\%} identical to RET-RGS1 (except its N-terminal extension) and 56{\%} identical to GAIP. Purified, recombinant RGSZ1, RET-RGS1, and GAIP each accelerated the hydrolysis of Gα(z)-GTP over 400-fold with K(m) values of ~2 nM. RGSZ1 was 100-fold selective for Gα(z) over Gα1, unusually specific among RGS proteins. Other enzymological properties of RGSZ1, brain G(z) GAP, and RET-RGS1 were identical; GAIP differed only in Mg2+ dependence and in its slightly lower selectivity for Gα(z). RGSZ1, RET-RGS1, and GAIP thus define a subfamily of G(z) GAPs within the RGS proteins. RGSZ1 has no obvious membrane-spanning region but is tightly membrane-bound in brain. Its regulatory activity in membranes depends on stable bilayer association. When co-reconstituted into phospholipid vesicles with G(z) and m2 muscarinic receptors, RGSZ1 increased agonist- stimulated GTPase >15-fold with EC50 <12 nM, but RGSZ1 added to the vesicle suspension was <0.1{\%} as active. RGSZ1, RET-RGS1, and GAIP share a cysteine string sequence, perhaps targeting them to secretory vesicles and allowing them to participate in the proposed control of secretion by G(z). Phosphorylation of Gα(z) by protein kinase C inhibited the GAP activity of RGSZ1 and other RGS proteins, providing a mechanism for potentiation of G(z) signaling by protein kinase C.",
author = "Jun Wang and Axel Ducret and Yaping Tu and Tohru Kozasa and Ruedi Aebersold and Ross, {Elliott M.}",
year = "1998",
month = "10",
day = "2",
doi = "10.1074/jbc.273.40.26014",
language = "English (US)",
volume = "273",
pages = "26014--26025",
journal = "Journal of Biological Chemistry",
issn = "0021-9258",
publisher = "American Society for Biochemistry and Molecular Biology Inc.",
number = "40",

}

TY - JOUR

T1 - RGSZ1, a G(z)-selective rgs protein in brain

T2 - Structure, membrane association, regulation by Gα(z) phosphorylation, and relationship to a G(z) gtpase-activating protein subfamily

AU - Wang, Jun

AU - Ducret, Axel

AU - Tu, Yaping

AU - Kozasa, Tohru

AU - Aebersold, Ruedi

AU - Ross, Elliott M.

PY - 1998/10/2

Y1 - 1998/10/2

N2 - We cloned the cDNA for human RGSZ1, the major G(z)-selective GTPase- activating protein (GAP) in brain (Wang, J., Tu, Y., Woodson, J., Song, X., and Ross, E. M. (1997) J. Biol. Chem. 272, 5732-5740) and a member of the RGS family of G protein GAPs. Its sequence is 83% identical to RET-RGS1 (except its N-terminal extension) and 56% identical to GAIP. Purified, recombinant RGSZ1, RET-RGS1, and GAIP each accelerated the hydrolysis of Gα(z)-GTP over 400-fold with K(m) values of ~2 nM. RGSZ1 was 100-fold selective for Gα(z) over Gα1, unusually specific among RGS proteins. Other enzymological properties of RGSZ1, brain G(z) GAP, and RET-RGS1 were identical; GAIP differed only in Mg2+ dependence and in its slightly lower selectivity for Gα(z). RGSZ1, RET-RGS1, and GAIP thus define a subfamily of G(z) GAPs within the RGS proteins. RGSZ1 has no obvious membrane-spanning region but is tightly membrane-bound in brain. Its regulatory activity in membranes depends on stable bilayer association. When co-reconstituted into phospholipid vesicles with G(z) and m2 muscarinic receptors, RGSZ1 increased agonist- stimulated GTPase >15-fold with EC50 <12 nM, but RGSZ1 added to the vesicle suspension was <0.1% as active. RGSZ1, RET-RGS1, and GAIP share a cysteine string sequence, perhaps targeting them to secretory vesicles and allowing them to participate in the proposed control of secretion by G(z). Phosphorylation of Gα(z) by protein kinase C inhibited the GAP activity of RGSZ1 and other RGS proteins, providing a mechanism for potentiation of G(z) signaling by protein kinase C.

AB - We cloned the cDNA for human RGSZ1, the major G(z)-selective GTPase- activating protein (GAP) in brain (Wang, J., Tu, Y., Woodson, J., Song, X., and Ross, E. M. (1997) J. Biol. Chem. 272, 5732-5740) and a member of the RGS family of G protein GAPs. Its sequence is 83% identical to RET-RGS1 (except its N-terminal extension) and 56% identical to GAIP. Purified, recombinant RGSZ1, RET-RGS1, and GAIP each accelerated the hydrolysis of Gα(z)-GTP over 400-fold with K(m) values of ~2 nM. RGSZ1 was 100-fold selective for Gα(z) over Gα1, unusually specific among RGS proteins. Other enzymological properties of RGSZ1, brain G(z) GAP, and RET-RGS1 were identical; GAIP differed only in Mg2+ dependence and in its slightly lower selectivity for Gα(z). RGSZ1, RET-RGS1, and GAIP thus define a subfamily of G(z) GAPs within the RGS proteins. RGSZ1 has no obvious membrane-spanning region but is tightly membrane-bound in brain. Its regulatory activity in membranes depends on stable bilayer association. When co-reconstituted into phospholipid vesicles with G(z) and m2 muscarinic receptors, RGSZ1 increased agonist- stimulated GTPase >15-fold with EC50 <12 nM, but RGSZ1 added to the vesicle suspension was <0.1% as active. RGSZ1, RET-RGS1, and GAIP share a cysteine string sequence, perhaps targeting them to secretory vesicles and allowing them to participate in the proposed control of secretion by G(z). Phosphorylation of Gα(z) by protein kinase C inhibited the GAP activity of RGSZ1 and other RGS proteins, providing a mechanism for potentiation of G(z) signaling by protein kinase C.

UR - http://www.scopus.com/inward/record.url?scp=0032475515&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0032475515&partnerID=8YFLogxK

U2 - 10.1074/jbc.273.40.26014

DO - 10.1074/jbc.273.40.26014

M3 - Article

C2 - 9748280

AN - SCOPUS:0032475515

VL - 273

SP - 26014

EP - 26025

JO - Journal of Biological Chemistry

JF - Journal of Biological Chemistry

SN - 0021-9258

IS - 40

ER -