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PMID: 18434502 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

The PIP2 binding mode of the C2 domains of rabphilin-3A.

Protein science : a publication of the Protein Society ·Vol. 17 ·No. 6 ·2008-06-00 ·Pages 1025-34

Montaville P, Coudevylle N, Radhakrishnan A, Leonov A, Zweckstetter M, Becker S

Abstract

Phosphatidylinositol-4,5-bisphosphate (PIP2) is a key player in the neurotransmitter release process. Rabphilin-3A is a neuronal C2 domain tandem containing protein that is involved in this process. Both its C2 domains (C2A and C2B) are able to bind PIP2. The investigation of the interactions of the two C2 domains with the PIP2 headgroup IP3 (inositol-1,4,5-trisphosphate) by NMR showed that a well-defined binding site can be described on the concave surface of each domain. The binding modes of the two domains are different. The binding of IP3 to the C2A domain is strongly enhanced by Ca(2+) and is characterized by a K(D) of 55 microM in the presence of a saturating concentration of Ca(2+) (5 mM). Reciprocally, the binding of IP3 increases the apparent Ca(2+)-binding affinity of the C2A domain in agreement with a Target-Activated Messenger Affinity (TAMA) mechanism. The C2B domain binds IP3 in a Ca(2+)-independent fashion with low affinity. These different PIP2 headgroup recognition modes suggest that PIP2 is a target of the C2A domain of rabphilin-3A while this phospholipid is an effector of the C2B domain.

MeSH Terms
Adaptor Proteins, Signal Transducing/chemistry,metabolism Animals Calcium/metabolism Models, Molecular Nerve Tissue Proteins/chemistry,metabolism Nuclear Magnetic Resonance, Biomolecular Phosphatidylinositol 4,5-Diphosphate/metabolism Phosphoserine/analogs & derivatives,metabolism Protein Binding Rats Vesicular Transport Proteins/chemistry,metabolism
Chemicals
Adaptor Proteins, Signal Transducing Nerve Tissue Proteins Phosphatidylinositol 4,5-Diphosphate Vesicular Transport Proteins rabphilin-3A Phosphoserine glycerophosphoserine Calcium
Authors & Affiliations
6 authors, click to expand affiliations / ORCID
Montaville Pierre
Department of NMR-based Structural Biology, Max-Planck-Institute for Biophysical Chemistry, 37077 Göttingen, Germany.
Coudevylle Nicolas
Radhakrishnan Anand
Leonov Andrei
Zweckstetter Markus
Becker Stefan
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Article Info
Journal
Protein science : a publication of the Protein Society
Abbr.
Protein Sci
ISSN
1469-896X
DOI
10.1110/ps.073326608
Published
2008-06-00
Epub
2008-00-23
Pages
1025-34
Language
English
Region
United States
NLM ID
9211750
PMCID
PMC2386734
Subset
IM
External Links
PubMed source DOI: 10.1110/ps.073326608 PMC Full Text View journal details
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