Insights Regarding Guanine Nucleotide Exchange from the Structure of a DENN-domain Protein Complexed with Its Rab GTPase Substrate

Overview

Journal Proc Natl Acad Sci U S A

Specialty Science

Date 2011 Nov 9

PMID 22065758

Citations 68

Authors

Xudong Wu

Michael J Bradley

Yiying Cai

Daniel Kummel

Enrique M De La Cruz

Francis A Barr

Karin M Reinisch

Affiliations

Soon will be listed here.

Abstract

Rab GTPases are key regulators of membrane traffic pathways within eukaryotic cells. They are specifically activated by guanine nucleotide exchange factors (GEFs), which convert them from their "inactive" GDP-bound form to the "active" GTP-bound form. In higher eukaryotes, proteins containing DENN-domains comprise a major GEF family. Here we describe at 2.1-Å resolution the first structure of a DENN-domain protein, DENND1B-S, complexed with its substrate Rab35, providing novel insights as to how DENN-domain GEFs interact with and activate Rabs. DENND1B-S is bi-lobed, and interactions with Rab35 are through conserved surfaces in both lobes. Rab35 binds via switch regions I and II, around the nucleotide-binding pocket. Positional shifts in Rab residues required for nucleotide binding may lower its affinity for bound GDP, and a conformational change in switch I, which makes the nucleotide-binding pocket more solvent accessible, likely also facilitates exchange.

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