title: Overexpression, purification, crystallization and preliminary X-ray crystallographic studies of a proline-specific aminopeptidase from Aneurinibacillus sp. strain AM-1.
abstract: To elucidate the structure and molecular mechanism of a characteristic proline-specific aminopeptidase produced by the thermophile Aneurinibacillus sp. strain AM-1, its gene was cloned and the recombinant protein was overexpressed in Escherichia coli, purified and crystallized using the hanging-drop vapour-diffusion method. X-ray diffraction data were collected to 1.8 A resolution from the recombinant aminopeptidase crystal. The crystals belong to the orthorhombic space group P2(1)2(1)2, with unit-cell parameters a = 93.62, b = 68.20, c = 76.84 A. A complete data set was also obtained from crystals of SeMet-substituted aminopeptidase. Data in the resolution range 20-2.1 A from the MAD data set from the SeMet-substituted crystal were used for phase determination.
authors.affiliation: Department of Applied Biochemistry, Kyoto Prefectural University, Shimogamo, Sakyo, Kyoto 606-8522, Japan.
authors.forename: Makoto
authors.initials: M
authors.lastname: Akioka
authors.forename: Hiroaki
authors.initials: H
authors.lastname: Nakano
authors.forename: Aya
authors.initials: A
authors.lastname: Horikiri
authors.forename: Yoshiyuki
authors.initials: Y
authors.lastname: Tsujimoto
authors.forename: Hiroshi
authors.initials: H
authors.lastname: Matsui
authors.forename: Tetsuya
authors.initials: T
authors.lastname: Shimizu
authors.forename: Toru
authors.initials: T
authors.lastname: Nakatsu
authors.forename: Hiroaki
authors.initials: H
authors.lastname: Kato
authors.forename: Kunihiko
authors.initials: K
authors.lastname: Watanabe
mesh_terms: D000626:Aminopeptidases / Q000096:biosynthesis* / Q000737:chemistry* / Q000302:isolation & purification; D001419:Bacteria / Q000201:enzymology; D003001:Cloning, Molecular; D003460:Crystallization; D018360:Crystallography, X-Ray; D004926:Escherichia coli / Q000201:enzymology; D011994:Recombinant Proteins / Q000737:chemistry
chemical_list: D011994:Recombinant Proteins; D000626:Aminopeptidases