Expression, purification, crystallization and preliminary X-ray diffraction studies of human liver regucalcin.
Regucalcin is a novel calcium ion (Ca(2+)) binding protein that does not contain an EF-hand motif as a Ca(2+)-binding domain and has been demonstrated to play a multi-functional role in many cell types. Human liver regucalcin, consisting of 299 amino-acid residues, was overexpressed in Escherichia coli, purified and crystallized by the vapour-diffusion method in the presence of polyethylene glycol 4000 as a precipitant. A native crystal diffracted to 2.8 A with synchrotron radiation and belongs to space group P2(1), with unit-cell parameters a = 64.87, b = 52.52, c = 86.38 A, beta = 99.86 degrees . Two molecules most probably exist in the asymmetric unit, corresponding to V(M) = 2.2 A(3) Da(-1). Heavy-atom derivative data were collected and the Pb derivative showed one high-occupancy site per molecule.
Warizaya
Masaichi
M
Exploratory Research Laboratory, Fujisawa Pharmaceutical Co. Ltd, 5-2-3 Tokodai, Tsukuba, Ibaraki 300-2698, Japan. masaichi_warizaya@po.fujisawa.co.jp
Kinoshita
Takayoshi
T
Yamaoka
Makiko
M
Shibata
Takashi
T
Saito
Noriko
N
Nakajima
Hidenori
H
Fujii
Takashi
T
D002135:Calcium-Binding Proteins / Q000737:chemistry* / Q000235:genetics / Q000302:isolation & purification*; D003460:Crystallization; D015870:Gene Expression; D005822:Genetic Vectors / Q000235:genetics; D006801:Humans; D047908:Intracellular Signaling Peptides and Proteins; D008099:Liver / Q000737:chemistry*; D015238:Sulfotransferases; D014961:X-Ray Diffraction
D002135:Calcium-Binding Proteins; D047908:Intracellular Signaling Peptides and Proteins; C513323:RGN protein, human; D015238:Sulfotransferases; C028152:alcohol sulfotransferase