Preliminary crystallographic analysis of the Escherichia coli YeaZ protein using the anomalous signal of a gadolinium derivative.
The Escherichia coli yeaZ gene encodes a 231-residue protein (Mr = 25,180) that belongs to a family of proteins that are conserved in various bacterial genomes. This protein of unknown function is predicted to be a hypothetical protease. The YeaZ protein was overexpressed in E. coli and crystallized at 298 K by the hanging-drop vapour-diffusion method. A MAD data set was collected using a gadolinium-derivative crystal that had been soaked with 0.1 M Gd-DOTMA. The data set contained data collected to a resolution of 2.7 A at two wavelengths at the L(III) absorption edge of gadolinium, while remote data were collected to a resolution of 2.28 A. The crystal belonged to the orthorhombic space group P2(1)2(1)2(1), with unit-cell parameters a = 76.3, b = 97.6, c = 141.9 A. Phasing using the MAD method confirmed there to be four monomers in the asymmetric unit related by two twofold axes as identified by the self-rotation function search.
Jeudy
Sandra
S
Information Génomique et Structurale, CNRS UPR 2589, IBSM, 31 Chemin Joseph Aiguier, 13402 Marseille Cedex 20, France.
Stelter
Meike
M
Coutard
Bruno
B
Kahn
Richard
R
Abergel
Chantal
C
D000595:Amino Acid Sequence; D003460:Crystallization; D018360:Crystallography, X-Ray; D003625:Data Collection; D004591:Electrophoresis, Polyacrylamide Gel; D004926:Escherichia coli / Q000737:chemistry*; D029968:Escherichia coli Proteins / Q000737:chemistry* / Q000302:isolation & purification; D005682:Gadolinium / Q000737:chemistry*; D015870:Gene Expression; D008969:Molecular Sequence Data; D016415:Sequence Alignment
D029968:Escherichia coli Proteins; C512523:YeaZ protein, E coli; D005682:Gadolinium