Open AccessOverexpression, purification, crystallization and preliminary X‐ray cystallographic studies of a proline‐specific aminopeptidase from Aneurinibacillus sp. strain AM‐1
Author(s) -
Akioka Makoto,
Nakano Hiroaki,
Horikiri Aya,
Tsujimoto Yoshiyuki,
Matsui Hiroshi,
Shimizu Tetsuya,
Nakatsu Toru,
Kato Hiroaki,
Watanabe Kunihiko
Publication year - 2006
Publication title -
acta crystallographica section f
Language(s) - English
Resource type - Journals
ISSN - 1744-3091
DOI - 10.1107/s1744309106047543
Subject(s) - orthorhombic crystal system , escherichia coli , aminopeptidase , crystallization , resolution (logic) , recombinant dna , strain (injury) , crystal (programming language) , leucine , thermophile , crystallography , microbiology and biotechnology , biochemistry , chemistry , biology , crystal structure , gene , amino acid , enzyme , organic chemistry , anatomy , artificial intelligence , computer science , programming language
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 Å resolution from the recombinant aminopeptidase crystal. The crystals belong to the orthorhombic space group P 2 1 2 1 2, with unit‐cell parameters a = 93.62, b = 68.20, c = 76.84 Å. A complete data set was also obtained from crystals of SeMet‐substituted aminopeptidase. Data in the resolution range 20–2.1 Å from the MAD data set from the SeMet‐substituted crystal were used for phase determination.