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Title: Cloning, expression, purification, crystallization and preliminary X-ray diffraction analysis of the glutamate-1-semialdehyde aminotransferase from Bacillus subtilis

Abstract

Crystals of glutamate-1-semialdehyde aminotransferase (GSAT) from B. subtilis were obtained and diffraction data were collected to 2.0 Å resolution. 5-Aminolevulinic acid (ALA) is the first committed universal precursor in the tetrapyrrole-biosynthesis pathway. Plants, algae and many other bacteria synthesize ALA from glutamate by a C5 pathway in which the carbon skeleton of glutamate is converted into ALA by a series of enzymes. Glutamate-1-semialdehyde aminotransferase (GSAT) is the last enzyme in this pathway. The gene that codes for GSAT was amplified from the cDNA library of Bacillus subtilis and overexpressed in Escherichia coli strain BL21(DE3). The protein was purified and crystallized. Well diffracting single crystals were obtained by the hanging-drop vapour-diffusion method. Preliminary X-ray diffraction studies yielded excellent diffraction data to a resolution of 2.0 Å.

Authors:
; ; ; ; ; ;  [1];  [2]
  1. Hefei National Laboratory for Physical Sciences at Microscale and School of Life Sciences, University of Science and Technology of China, 96 Jinzhai Road, Hefei, Anhui 230027 (China)
  2. (China)
Publication Date:
OSTI Identifier:
22356321
Resource Type:
Journal Article
Resource Relation:
Journal Name: Acta Crystallographica. Section F; Journal Volume: 62; Journal Issue: Pt 5; Other Information: PMCID: PMC2219978; PMID: 16682782; PUBLISHER-ID: ll5059; OAI: oai:pubmedcentral.nih.gov:2219978; Copyright (c) International Union of Crystallography 2006; Country of input: International Atomic Energy Agency (IAEA)
Country of Publication:
United Kingdom
Language:
English
Subject:
75 CONDENSED MATTER PHYSICS, SUPERCONDUCTIVITY AND SUPERFLUIDITY; CARBON; CRYSTALLIZATION; DIFFUSION; ESCHERICHIA COLI; MONOCRYSTALS; PRECURSOR; RESOLUTION; STRAINS; X-RAY DIFFRACTION

Citation Formats

Lv, Xinhuai, Fan, Jun, Ge, Honghua, Gao, Yongxiang, Zhang, Xiao, Teng, Maikun, E-mail: mkteng@ustc.edu.cn, Niu, Liwen, E-mail: mkteng@ustc.edu.cn, and Key Laboratory of Structural Biology, Chinese Academy of Sciences, 96 Jinzhai Road, Hefei, Anhui 230027. Cloning, expression, purification, crystallization and preliminary X-ray diffraction analysis of the glutamate-1-semialdehyde aminotransferase from Bacillus subtilis. United Kingdom: N. p., 2006. Web. doi:10.1107/S1744309106013121.
Lv, Xinhuai, Fan, Jun, Ge, Honghua, Gao, Yongxiang, Zhang, Xiao, Teng, Maikun, E-mail: mkteng@ustc.edu.cn, Niu, Liwen, E-mail: mkteng@ustc.edu.cn, & Key Laboratory of Structural Biology, Chinese Academy of Sciences, 96 Jinzhai Road, Hefei, Anhui 230027. Cloning, expression, purification, crystallization and preliminary X-ray diffraction analysis of the glutamate-1-semialdehyde aminotransferase from Bacillus subtilis. United Kingdom. doi:10.1107/S1744309106013121.
Lv, Xinhuai, Fan, Jun, Ge, Honghua, Gao, Yongxiang, Zhang, Xiao, Teng, Maikun, E-mail: mkteng@ustc.edu.cn, Niu, Liwen, E-mail: mkteng@ustc.edu.cn, and Key Laboratory of Structural Biology, Chinese Academy of Sciences, 96 Jinzhai Road, Hefei, Anhui 230027. Mon . "Cloning, expression, purification, crystallization and preliminary X-ray diffraction analysis of the glutamate-1-semialdehyde aminotransferase from Bacillus subtilis". United Kingdom. doi:10.1107/S1744309106013121.
@article{osti_22356321,
title = {Cloning, expression, purification, crystallization and preliminary X-ray diffraction analysis of the glutamate-1-semialdehyde aminotransferase from Bacillus subtilis},
author = {Lv, Xinhuai and Fan, Jun and Ge, Honghua and Gao, Yongxiang and Zhang, Xiao and Teng, Maikun, E-mail: mkteng@ustc.edu.cn and Niu, Liwen, E-mail: mkteng@ustc.edu.cn and Key Laboratory of Structural Biology, Chinese Academy of Sciences, 96 Jinzhai Road, Hefei, Anhui 230027},
abstractNote = {Crystals of glutamate-1-semialdehyde aminotransferase (GSAT) from B. subtilis were obtained and diffraction data were collected to 2.0 Å resolution. 5-Aminolevulinic acid (ALA) is the first committed universal precursor in the tetrapyrrole-biosynthesis pathway. Plants, algae and many other bacteria synthesize ALA from glutamate by a C5 pathway in which the carbon skeleton of glutamate is converted into ALA by a series of enzymes. Glutamate-1-semialdehyde aminotransferase (GSAT) is the last enzyme in this pathway. The gene that codes for GSAT was amplified from the cDNA library of Bacillus subtilis and overexpressed in Escherichia coli strain BL21(DE3). The protein was purified and crystallized. Well diffracting single crystals were obtained by the hanging-drop vapour-diffusion method. Preliminary X-ray diffraction studies yielded excellent diffraction data to a resolution of 2.0 Å.},
doi = {10.1107/S1744309106013121},
journal = {Acta Crystallographica. Section F},
number = Pt 5,
volume = 62,
place = {United Kingdom},
year = {Mon May 01 00:00:00 EDT 2006},
month = {Mon May 01 00:00:00 EDT 2006}
}