Open Access
Cloning, expression, purification, crystallization and preliminary X‐ray diffraction crystallographic study of human synaptotagmin 5 C2A domain
Author(s) -
Qiu Xiaoting,
Huang Kai,
Liu Yiwei,
Zhang Xiao,
Gao Yongxiang
Publication year - 2011
Publication title -
acta crystallographica section f
Language(s) - English
Resource type - Journals
ISSN - 1744-3091
DOI - 10.1107/s1744309111032155
Subject(s) - synaptotagmin 1 , crystallography , chemistry , vesicle fusion , exocytosis , crystallization , microbiology and biotechnology , biology , biochemistry , vesicle , synaptic vesicle , secretion , membrane , organic chemistry
Synaptotagmin acts as the Ca 2+ sensor for neural and endocrine exocytosis. Synaptotagmin 5 has been demonstrated to play a key role in the acquisition of cathepsin D and the vesicular proton ATPase and in Ca 2+ ‐dependent insulin exocytosis. The C2 domains modulate the interaction of synaptotagmin with the phospholipid bilayer of the presynaptic terminus and effector proteins such as the SNARE complex. This study reports the cloning, expression in Escherichia coli , purification, crystallization and preliminary X‐ray analysis of the C2A domain of human synaptotagmin 5 with an N‐terminal His 6 tag. The crystals diffracted to 1.90 Å resolution and belonged to the hexagonal space group P 6 5 , with unit‐cell parameters a = b = 93.97, c = 28.05 Å. A preliminary model of the protein structure has been built and refinement of the model is ongoing.