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PMID: 24302762 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Structural insights into the Ca2+ and PI(4,5)P2 binding modes of the C2 domains of rabphilin 3A and synaptotagmin 1.

Proceedings of the National Academy of Sciences of the United States of America ·Vol. 110 ·No. 51 ·2013-12-17 ·Pages 20503-8

Guillén J, Ferrer-Orta C, Buxaderas M, Pérez-Sánchez D, Guerrero-Valero M, Luengo-Gil G, Pous J, Guerra P, Gómez-Fernández JC, Verdaguer N, Corbalán-García S

Abstract

Proteins containing C2 domains are the sensors for Ca(2+) and PI(4,5)P2 in a myriad of secretory pathways. Here, the use of a free-mounting system has enabled us to capture an intermediate state of Ca(2+) binding to the C2A domain of rabphilin 3A that suggests a different mechanism of ion interaction. We have also determined the structure of this domain in complex with PI(4,5)P2 and IP3 at resolutions of 1.75 and 1.9 Å, respectively, unveiling that the polybasic cluster formed by strands β3-β4 is involved in the interaction with the phosphoinositides. A comparative study demonstrates that the C2A domain is highly specific for PI(4,5)P2/PI(3,4,5)P3, whereas the C2B domain cannot discriminate among any of the diphosphorylated forms. Structural comparisons between C2A domains of rabphilin 3A and synaptotagmin 1 indicated the presence of a key glutamic residue in the polybasic cluster of synaptotagmin 1 that abolishes the interaction with PI(4,5)P2. Together, these results provide a structural explanation for the ability of different C2 domains to pull plasma and vesicle membranes close together in a Ca(2+)-dependent manner and reveal how this family of proteins can use subtle structural changes to modulate their sensitivity and specificity to various cellular signals.

Keywords
PIP2 calcium vesicle fusion
MeSH Terms
Adaptor Proteins, Signal Transducing/chemistry,genetics,metabolism Calcium/chemistry,metabolism Cell Membrane/chemistry,genetics,metabolism Crystallography, X-Ray Humans Nerve Tissue Proteins/chemistry,genetics,metabolism Phosphatidylinositol 4,5-Diphosphate/chemistry,genetics,metabolism Protein Structure, Tertiary Structure-Activity Relationship Synaptotagmin I/chemistry,genetics,metabolism Vesicular Transport Proteins/chemistry,genetics,metabolism
Chemicals
Adaptor Proteins, Signal Transducing Nerve Tissue Proteins Phosphatidylinositol 4,5-Diphosphate Synaptotagmin I Vesicular Transport Proteins rabphilin-3A Calcium
Authors & Affiliations
11 authors, click to expand affiliations / ORCID
Guillén Jaime
Departamento de Bioquímica y Biología Molecular A, Facultad de Veterinaria, Regional Campus of International Excellence "Campus Mare Nostrum," Universidad de Murcia, 30100 Murcia, Spain.
Ferrer-Orta Cristina
Buxaderas Mònica
Pérez-Sánchez Dolores
Guerrero-Valero Marta
Luengo-Gil Ginés
Pous Joan
Guerra Pablo
Gómez-Fernández Juan C
Verdaguer Nuria
Corbalán-García Senena
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Article Info
Journal
Proceedings of the National Academy of Sciences of the United States of America
Abbr.
Proc Natl Acad Sci U S A
ISSN
1091-6490
DOI
10.1073/pnas.1316179110
Published
2013-12-17
Epub
2013-00-03
Pages
20503-8
Language
English
Region
United States
NLM ID
7505876
PMCID
PMC3870689
Subset
IM
Databases
PDB
4LT7, 4NP9, 4NS0
External Links
PubMed source DOI: 10.1073/pnas.1316179110 PMC Full Text View journal details
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