title: Cloning, expression, purification, crystallization and preliminary X-ray diffraction crystallographic study of human synaptotagmin 5 C2A domain.
abstract: 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 P6(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.
authors.affiliation: Hefei National Laboratory for Physical Sciences at Microscale, School of Life Sciences, University of Science and Technology of China, 96 Jinzhai Road, Hefei, Anhui 230026, People's Republic of China.
authors.forename: Xiaoting
authors.initials: X
authors.lastname: Qiu
authors.forename: Kai
authors.initials: K
authors.lastname: Huang
authors.forename: Yiwei
authors.initials: Y
authors.lastname: Liu
authors.forename: Xiao
authors.initials: X
authors.lastname: Zhang
authors.forename: Yongxiang
authors.initials: Y
authors.lastname: Gao
mesh_terms: D003001:Cloning, Molecular; D003460:Crystallization; D018360:Crystallography, X-Ray; D015870:Gene Expression; D006801:Humans; D015394:Molecular Structure; D050857:Synaptotagmins / Q000737:chemistry* / Q000235:genetics / Q000302:isolation & purification
chemical_list: D050857:Synaptotagmins
references.citation: Cell. 1995 Mar 24;80(6):929-38
references.pmid: 7697723
references.citation: Nat Rev Mol Cell Biol. 2002 Jul;3(7):498-508
references.pmid: 12094216
references.citation: Acta Crystallogr D Biol Crystallogr. 2011 Apr;67(Pt 4):355-67
references.pmid: 21460454
references.citation: Proc Natl Acad Sci U S A. 2005 Apr 5;102(14):5210-4
references.pmid: 15793006
references.citation: Methods Enzymol. 1997;276:307-26
references.pmid: 27754618
references.citation: Science. 1986 Aug 22;233(4766):853-9
references.pmid: 3755547
references.citation: PLoS One. 2010 Sep 02;5(9):
references.pmid: 20824061
references.citation: Neuron. 1997 Jan;18(1):133-42
references.pmid: 9010211
references.citation: J Biol Chem. 2002 Mar 8;277(10):7629-32
references.pmid: 11739399
references.citation: Protein Sci. 2004 Oct;13(10):2665-72
references.pmid: 15340165
references.citation: Annu Rev Neurosci. 2004;27:509-47
references.pmid: 15217342
references.citation: Cell Calcium. 1986 Dec;7(5-6):339-52
references.pmid: 3545486
references.citation: J Biol Chem. 2002 Jul 5;277(27):24499-505
references.pmid: 12006594
references.citation: Acta Crystallogr D Biol Crystallogr. 2010 Jan;66(Pt 1):22-5
references.pmid: 20057045
references.citation: PLoS Pathog. 2009 Oct;5(10):e1000628
references.pmid: 19834555
references.citation: Monogr Neural Sci. 1980;7:106-16
references.pmid: 6112701
references.citation: Nat Struct Mol Biol. 2010 Mar;17(3):325-31
references.pmid: 20173762
references.citation: Biochemistry. 2007 Nov 13;46(45):13041-8
references.pmid: 17956130
references.citation: Fed Proc. 1982 May;41(7):2265-72
references.pmid: 6122609
references.citation: J Cell Biol. 1999 Nov 1;147(3):589-98
references.pmid: 10545502
references.citation: Acta Crystallogr D Biol Crystallogr. 2004 Dec;60(Pt 12 Pt 1):2126-32
references.pmid: 15572765
references.citation: Biophys J. 2005 Jul;89(1):690-702
references.pmid: 15821166
references.citation: J Mol Biol. 1968 Apr 28;33(2):491-7
references.pmid: 5700707
references.citation: J Cell Sci. 2004 Jul 1;117(Pt 15):3119-27
references.pmid: 15190121