PDBsum entry 4lcv

Metal binding protein

PDB id: 4lcv

Contents

Protein chains

123 a.a.

131 a.a.

Ligands

SO4 ×4

BME

FLC

Metals

_CA ×2

Waters ×261

PDB id:

4lcv

Name:

Metal binding protein

Title:

Crystal structure of doc2b c2a domain

Structure:

Double c2-like domain-containing protein beta. Chain: a, b, c, d. Fragment: c2a domain (unp residues 125-255). Synonym: doc2-beta. Engineered: yes

Source:

Rattus norvegicus. Brown rat,rat,rats. Organism_taxid: 10116. Gene: doc2b. Expressed in: escherichia coli. Expression_system_taxid: 562

Resolution:

2.00Å R-factor: 0.189 R-free: 0.232

Authors:

M.Giladi,L.Almagor,J.A.Hirsch

Key ref:

M.Giladi et al. (2013). The C2B domain is the primary Ca2+ sensor in DOC2B: a structural and functional analysis. J Mol Biol, 425, 4629-4641. PubMed id: 23994332 DOI: 10.1016/j.jmb.2013.08.017

Date:

23-Jun-13 Release date: 11-Sep-13

Protein chain

P70610  (DOC2B_RAT) -  Double C2-like domain-containing protein beta from Rattus norvegicus

Seq: 412 a.a.

Struc: 123 a.a.

Protein chains

P70610  (DOC2B_RAT) -  Double C2-like domain-containing protein beta from Rattus norvegicus

Seq: 412 a.a.

Struc: 131 a.a.*

* PDB and UniProt seqs differ at 1 residue position (black cross)

DOI no: 10.1016/j.jmb.2013.08.017

J Mol Biol 425:4629-4641 (2013)

PubMed id: 23994332

The C2B domain is the primary Ca2+ sensor in DOC2B: a structural and functional analysis.

M.Giladi, L.Michaeli, L.Almagor, D.Bar-On, T.Buki, U.Ashery, D.Khananshvili, J.A.Hirsch.

ABSTRACT

DOC2B (double-C2 domain) protein is thought to be a high-affinity Ca(2+) sensor for spontaneous and asynchronous neurotransmitter release. To elucidate the molecular features underlying its physiological role, we determined the crystal structures of its isolated C2A and C2B domains and examined their Ca(2+)-binding properties. We further characterized the solution structure of the tandem domains (C2AB) using small-angle X-ray scattering. In parallel, we tested structure-function correlates with live cell imaging tools. We found that, despite striking structural similarity, C2B binds Ca(2+) with considerably higher affinity than C2A. The C2AB solution structure is best modeled as two domains with a highly flexible orientation and no difference in the presence or absence of Ca(2+). In addition, kinetic studies of C2AB demonstrate that, in the presence of unilamellar vesicles, Ca(2+) binding is stabilized, as reflected by the ~10-fold slower rate of Ca(2+) dissociation than in the absence of vesicles. In cells, isolated C2B translocates to the plasma membrane (PM) with an EC50 of 400nM while the C2A does not translocate at submicromolar Ca(2+) concentrations, supporting the biochemical observations. Nevertheless, C2AB translocates to the PM with an ~2-fold lower EC50 and to a greater extent than C2B. Our results, together with previous studies, reveal that the C2B is the primary Ca(2+) sensing unit in DOC2B, whereas C2A enhances the interaction of C2AB with the PM.