English
 
User Manual Privacy Policy Disclaimer Contact us
  Advanced SearchBrowse

Item

ITEM ACTIONSEXPORT
  Structural insights into the Calmodulin−Munc13 interaction obtained by cross-linking and mass spectrometry

Dimova, K., Kalkhof, S., Pottratz, I., Ihling, C., Rodriguez-Castaneda, F., Liepold, T., et al. (2009). Structural insights into the Calmodulin−Munc13 interaction obtained by cross-linking and mass spectrometry. Biochemistry, 48: 10.1021/bi900300r, pp. 5908-5921. Retrieved from http://pubs.acs.org/doi/full/10.1021/bi900300r?prevSearch=&searchHistoryKey=.

Item is

Basic

show hide
Item Permalink: http://hdl.handle.net/11858/00-001M-0000-0012-D846-A Version Permalink: http://hdl.handle.net/11858/00-001M-0000-0027-EF95-B
Genre: Journal Article

Files

show Files

Locators

show

Creators

show
hide
 Creators:
Dimova, K.1, Author              
Kalkhof, S., Author
Pottratz, I., Author
Ihling, C., Author
Rodriguez-Castaneda, F.2, Author              
Liepold, T., Author
Griesinger, C.3, Author
Brose, N., Author
Sinz, A., Author
Jahn, O., Author
Affiliations:
1Department of Molecular Biology, MPI for biophysical chemistry, Max Planck Society, ou_578628              
2Department of NMR Based Structural Biology, MPI for biophysical chemistry, Max Planck Society, ou_578567              
3Max Planck Society, ou_persistent13              

Content

show
hide
Free keywords: -
 Abstract: Munc13 proteins are essential regulators of synaptic vesicle priming and play a key role in adaptive synaptic plasticity phenomena. We recently identified and characterized the Ca2+-dependent interaction of Munc13 and calmodulin (CaM) as the molecular mechanism linking changes in residual Ca2+ concentrations to presynaptic vesicle priming and short-term plasticity. Here, we used peptidic photoprobes covering the established CaM-binding motif of Munc13 for photoaffinity labeling (PAL) of CaM, followed by structural characterization of the covalent photoadducts. Our innovative analytical workflow based on isotopically labeled CaM and mass spectrometry revealed that, in the bound state, the hydrophobic anchor residue of the CaM-binding motif in Munc13s contacts two distinct methionine residues in the C-terminal domain of CaM. To address the orientation of the peptide during binding, we obtained additional distance constraints from the mass spectrometric analysis of chemically cross-linked CaM−Munc13 peptide adducts. The constraints from both complementary cross-linking approaches were integrated into low-resolution three-dimensional structure models of the CaM−Munc13 peptide complexes. Our experimental data are best compatible with the structure of the complex formed by CaM and a CaM-binding peptide derived from neuronal NO synthase and show that Munc13−1 and ubMunc13−2 bind to CaM in an antiparallel orientation through a 1-5-8 motif. The structural information about the CaM−Munc13 peptide complexes will facilitate the design of Munc13 variants with altered CaM affinity and thereby advance the detailed functional analysis of the role of Munc13 proteins in synaptic transmission and plasticity.

Details

show
hide
Language(s): eng - English
 Dates: 2009-05-19
 Publication Status: Published in print
 Pages: -
 Publishing info: -
 Table of Contents: -
 Rev. Method: -
 Degree: -

Event

show

Legal Case

show

Project information

show

Source 1

show
hide
Title: Biochemistry
Source Genre: Journal
 Creator(s):
Affiliations:
Publ. Info: -
Pages: - Volume / Issue: 48 Sequence Number: 10.1021/bi900300r Start / End Page: 5908 - 5921 Identifier: -