Recognition of the Activated States of Gα13 by the rgRGS Domain of PDZRhoGEF

Zhe Chen; William D. Singer; Shahab M. Danesh; Paul C. Sternweis; Stephen R. Sprang

doi:10.1016/j.str.2008.07.009

Recognition of the Activated States of Gα13 by the rgRGS Domain of PDZRhoGEF

Zhe Chen
, William D. Singer
, Shahab M. Danesh
, Paul C. Sternweis
, Stephen R. Sprang

Division of Biological and Biomedical Sciences

University of Texas Southwestern Medical Center

Research output: Contribution to journal › Article › peer-review

42 Scopus citations

Abstract

G12 class heterotrimeric G proteins stimulate RhoA activation by RGS-RhoGEFs. However, p115RhoGEF is a GTPase Activating Protein (GAP) toward Gα13, whereas PDZRhoGEF is not. We have characterized the interaction between the PDZRhoGEF rgRGS domain (PRG-rgRGS) and the alpha subunit of G13 and have determined crystal structures of their complexes in both the inactive state bound to GDP and the active states bound to GDP•AlF (transition state) and GTPγS (Michaelis complex). PRG-rgRGS interacts extensively with the helical domain and the effector-binding sites on Gα13 through contacts that are largely conserved in all three nucleotide-bound states, although PRG-rgRGS has highest affinity to the Michaelis complex. An acidic motif in the N terminus of PRG-rgRGS occupies the GAP binding site of Gα13 and is flexible in the GDP•AlF complex but well ordered in the GTPγS complex. Replacement of key residues in this motif with their counterparts in p115RhoGEF confers GAP activity.

Original language	English
Pages (from-to)	1532-1543
Number of pages	12
Journal	Structure
Volume	16
Issue number	10
DOIs	https://doi.org/10.1016/j.str.2008.07.009
State	Published - Oct 8 2008

Funding

We thank Jana Hadas, Helen Aronovich, Celestine Thomas, and Stephen Gutowski for technical assistance, and Chad Brautigam and the staff of APS for assistance with data collection. This work was supported by National Institute of Health Grants GM31954 (to P.C.S.), and DK46371 (to S.R.S.), by the Robert A. Welch foundation (I-1262 to P.C.S. and I-1229 S.R.S.), the Alfred and Mabel Gilman Chair in Molecular Pharmacology (P.C.S.), and the John W. and Rhonda K. Pate Professorship in Biochemistry (to S.R.S.).

Funder number
GM31954
I-1262, I-1229
R01DK046371

Keywords

MICROBIO
PROTEINS

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10.1016/j.str.2008.07.009

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title = "Recognition of the Activated States of Gα13 by the rgRGS Domain of PDZRhoGEF",

abstract = "G12 class heterotrimeric G proteins stimulate RhoA activation by RGS-RhoGEFs. However, p115RhoGEF is a GTPase Activating Protein (GAP) toward Gα13, whereas PDZRhoGEF is not. We have characterized the interaction between the PDZRhoGEF rgRGS domain (PRG-rgRGS) and the alpha subunit of G13 and have determined crystal structures of their complexes in both the inactive state bound to GDP and the active states bound to GDP•AlF (transition state) and GTPγS (Michaelis complex). PRG-rgRGS interacts extensively with the helical domain and the effector-binding sites on Gα13 through contacts that are largely conserved in all three nucleotide-bound states, although PRG-rgRGS has highest affinity to the Michaelis complex. An acidic motif in the N terminus of PRG-rgRGS occupies the GAP binding site of Gα13 and is flexible in the GDP•AlF complex but well ordered in the GTPγS complex. Replacement of key residues in this motif with their counterparts in p115RhoGEF confers GAP activity.",

keywords = "MICROBIO, PROTEINS",

author = "Zhe Chen and Singer, \{William D.\} and Danesh, \{Shahab M.\} and Sternweis, \{Paul C.\} and Sprang, \{Stephen R.\}",

note = "Funding Information: We thank Jana Hadas, Helen Aronovich, Celestine Thomas, and Stephen Gutowski for technical assistance, and Chad Brautigam and the staff of APS for assistance with data collection. This work was supported by National Institute of Health Grants GM31954 (to P.C.S.), and DK46371 (to S.R.S.), by the Robert A. Welch foundation (I-1262 to P.C.S. and I-1229 S.R.S.), the Alfred and Mabel Gilman Chair in Molecular Pharmacology (P.C.S.), and the John W. and Rhonda K. Pate Professorship in Biochemistry (to S.R.S.). ",

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doi = "10.1016/j.str.2008.07.009",

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AU - Chen, Zhe

AU - Singer, William D.

AU - Danesh, Shahab M.

AU - Sternweis, Paul C.

AU - Sprang, Stephen R.

N1 - Funding Information: We thank Jana Hadas, Helen Aronovich, Celestine Thomas, and Stephen Gutowski for technical assistance, and Chad Brautigam and the staff of APS for assistance with data collection. This work was supported by National Institute of Health Grants GM31954 (to P.C.S.), and DK46371 (to S.R.S.), by the Robert A. Welch foundation (I-1262 to P.C.S. and I-1229 S.R.S.), the Alfred and Mabel Gilman Chair in Molecular Pharmacology (P.C.S.), and the John W. and Rhonda K. Pate Professorship in Biochemistry (to S.R.S.).

PY - 2008/10/8

Y1 - 2008/10/8

N2 - G12 class heterotrimeric G proteins stimulate RhoA activation by RGS-RhoGEFs. However, p115RhoGEF is a GTPase Activating Protein (GAP) toward Gα13, whereas PDZRhoGEF is not. We have characterized the interaction between the PDZRhoGEF rgRGS domain (PRG-rgRGS) and the alpha subunit of G13 and have determined crystal structures of their complexes in both the inactive state bound to GDP and the active states bound to GDP•AlF (transition state) and GTPγS (Michaelis complex). PRG-rgRGS interacts extensively with the helical domain and the effector-binding sites on Gα13 through contacts that are largely conserved in all three nucleotide-bound states, although PRG-rgRGS has highest affinity to the Michaelis complex. An acidic motif in the N terminus of PRG-rgRGS occupies the GAP binding site of Gα13 and is flexible in the GDP•AlF complex but well ordered in the GTPγS complex. Replacement of key residues in this motif with their counterparts in p115RhoGEF confers GAP activity.

AB - G12 class heterotrimeric G proteins stimulate RhoA activation by RGS-RhoGEFs. However, p115RhoGEF is a GTPase Activating Protein (GAP) toward Gα13, whereas PDZRhoGEF is not. We have characterized the interaction between the PDZRhoGEF rgRGS domain (PRG-rgRGS) and the alpha subunit of G13 and have determined crystal structures of their complexes in both the inactive state bound to GDP and the active states bound to GDP•AlF (transition state) and GTPγS (Michaelis complex). PRG-rgRGS interacts extensively with the helical domain and the effector-binding sites on Gα13 through contacts that are largely conserved in all three nucleotide-bound states, although PRG-rgRGS has highest affinity to the Michaelis complex. An acidic motif in the N terminus of PRG-rgRGS occupies the GAP binding site of Gα13 and is flexible in the GDP•AlF complex but well ordered in the GTPγS complex. Replacement of key residues in this motif with their counterparts in p115RhoGEF confers GAP activity.

KW - MICROBIO

KW - PROTEINS

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JO - Structure

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ER -

Recognition of the Activated States of Gα13 by the rgRGS Domain of PDZRhoGEF

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