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Title: Crystallization and preliminary X-ray diffraction of the Munc18c–syntaxin4{sub 1–29} complex

Abstract

Cocrystallization with a peptide, free-interface diffusion crystal chips and crystal dehydration were important in the production of diffraction-quality crystals of the Munc18c protein that helps to regulate membrane fusion. The production of diffraction-quality crystals of Munc18c, a protein involved in regulating vesicular exocytosis in mammals, is reported. The diffraction resolution of Munc18c crystals was optimized by (i) cocrystallizing with a peptide fragment of the Munc18c functional binding partner syntaxin4, (ii) using nanolitre free-interface diffusion crystallization-screening chips and microlitre hanging-drop vapour diffusion and (iii) applying a post-crystallization dehydration treatment. Crystals belonging to the cubic space group P2{sub 1}3, with unit-cell parameters a = b = c = 170.8 Å, α = β = γ = 90°, were generated that diffract to 3.7 Å resolution on a laboratory X-ray source.

Authors:
; ; ; ;  [1];  [2];  [1]
  1. Institute for Molecular Bioscience and Special Research Centre for Functional and Applied Genomics, The University of Queensland, Brisbane, QLD 4072 (Australia)
  2. Garvan Institute of Medical Research, Darlinghurst, NSW 2010 (Australia)
Publication Date:
OSTI Identifier:
22360340
Resource Type:
Journal Article
Resource Relation:
Journal Name: Acta Crystallographica. Section F; Journal Volume: 63; Journal Issue: Pt 6; Other Information: PMCID: PMC2335071; PMID: 17554178; PUBLISHER-ID: bo5021; OAI: oai:pubmedcentral.nih.gov:2335071; Copyright (c) International Union of Crystallography 2007; Country of input: International Atomic Energy Agency (IAEA)
Country of Publication:
United Kingdom
Language:
English
Subject:
75 CONDENSED MATTER PHYSICS, SUPERCONDUCTIVITY AND SUPERFLUIDITY; CRYSTALLIZATION; CRYSTALS; DEHYDRATION; DIFFUSION; INTERFACES; MEMBRANES; RESOLUTION; SCREENING; SPACE GROUPS; X-RAY DIFFRACTION; X-RAY SOURCES

Citation Formats

Latham, Catherine F., Hu, Shu-Hong, Gee, Christine L., Armishaw, Chris J., Alewood, Paul F., James, David E., and Martin, Jennifer L., E-mail: j.martin@imb.uq.edu.au. Crystallization and preliminary X-ray diffraction of the Munc18c–syntaxin4{sub 1–29} complex. United Kingdom: N. p., 2007. Web. doi:10.1107/S1744309107022361.
Latham, Catherine F., Hu, Shu-Hong, Gee, Christine L., Armishaw, Chris J., Alewood, Paul F., James, David E., & Martin, Jennifer L., E-mail: j.martin@imb.uq.edu.au. Crystallization and preliminary X-ray diffraction of the Munc18c–syntaxin4{sub 1–29} complex. United Kingdom. doi:10.1107/S1744309107022361.
Latham, Catherine F., Hu, Shu-Hong, Gee, Christine L., Armishaw, Chris J., Alewood, Paul F., James, David E., and Martin, Jennifer L., E-mail: j.martin@imb.uq.edu.au. Fri . "Crystallization and preliminary X-ray diffraction of the Munc18c–syntaxin4{sub 1–29} complex". United Kingdom. doi:10.1107/S1744309107022361.
@article{osti_22360340,
title = {Crystallization and preliminary X-ray diffraction of the Munc18c–syntaxin4{sub 1–29} complex},
author = {Latham, Catherine F. and Hu, Shu-Hong and Gee, Christine L. and Armishaw, Chris J. and Alewood, Paul F. and James, David E. and Martin, Jennifer L., E-mail: j.martin@imb.uq.edu.au},
abstractNote = {Cocrystallization with a peptide, free-interface diffusion crystal chips and crystal dehydration were important in the production of diffraction-quality crystals of the Munc18c protein that helps to regulate membrane fusion. The production of diffraction-quality crystals of Munc18c, a protein involved in regulating vesicular exocytosis in mammals, is reported. The diffraction resolution of Munc18c crystals was optimized by (i) cocrystallizing with a peptide fragment of the Munc18c functional binding partner syntaxin4, (ii) using nanolitre free-interface diffusion crystallization-screening chips and microlitre hanging-drop vapour diffusion and (iii) applying a post-crystallization dehydration treatment. Crystals belonging to the cubic space group P2{sub 1}3, with unit-cell parameters a = b = c = 170.8 Å, α = β = γ = 90°, were generated that diffract to 3.7 Å resolution on a laboratory X-ray source.},
doi = {10.1107/S1744309107022361},
journal = {Acta Crystallographica. Section F},
number = Pt 6,
volume = 63,
place = {United Kingdom},
year = {Fri Jun 01 00:00:00 EDT 2007},
month = {Fri Jun 01 00:00:00 EDT 2007}
}
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