Expression, purification, crystallization and preliminary diffraction studies of the mammalian DAG kinase homologue YegS from Escherichia coli.
yegS is a gene encoding a 32 kDa cytosolic protein with unknown function but with strong sequence homology to a family of structurally uncharacterized eukaryotic non-protein kinases: diacylglycerol kinases, sphingosine kinases and ceramide kinases. Here, the overexpression, crystallization and preliminary diffraction analysis of Escherichia coli YegS are reported. The crystals belong to space group P2(1), with unit-cell parameters a = 42.4, b = 166.1, c = 48.5 A, beta = 96.97 degrees. The presence of a dimer in the asymmetric unit was estimated to give a Matthews coefficient (VM) of 2.5 A3 Da(-1) and a solvent content of 50.8%(v/v). Single-wavelength diffraction data were collected to a resolution of 1.9 A using synchrotron radiation.
Bakali
M Amin
MA
Department of Biochemistry and Biophysics, Stockholm University, SE-10691 Stockholm, Sweden.
Nordlund
Pär
P
Hallberg
B Martin
BM
D003460:Crystallization / Q000379:methods; D018360:Crystallography, X-Ray; D019852:Diacylglycerol Kinase / Q000737:chemistry* / Q000302:isolation & purification; D004926:Escherichia coli / Q000737:chemistry* / Q000378:metabolism; D029968:Escherichia coli Proteins / Q000096:biosynthesis / Q000737:chemistry* / Q000302:isolation & purification
D029968:Escherichia coli Proteins; D019852:Diacylglycerol Kinase