Enzymatic characterization and crystal structure of biosynthetic alanine racemase from Pseudomonas aeruginosa PAO1

被引:3
作者
Dong, Hui [1 ,2 ]
Han, Qingqing [3 ]
Guo, Yu [4 ,5 ,6 ]
Ju, Jiansong [3 ]
Wang, Shanshan [4 ]
Yuan, Chao [4 ,5 ,6 ]
Long, Wei [1 ,2 ]
He, Xin [1 ,2 ]
Xu, Shujing [3 ]
Li, Sheng [4 ]
机构
[1] Chinese Acad Med Sci, Inst Radiat Med, Key Lab Tianjin Radiat & Mol Nucl Med, Tianjin 300192, Peoples R China
[2] Peking Union Med Coll, Tianjin 300192, Peoples R China
[3] Hebei Normal Univ, Coll Life Sci, Shijiazhuang 050024, Hebei, Peoples R China
[4] ShanghaiTech Univ, Shanghai Inst Adv Immunochem Studies, Shanghai 201210, Peoples R China
[5] Univ Chinese Acad Sci, Beijing 100049, Peoples R China
[6] ShanghaiTech Univ, Sch Life Sci & Technol, Shanghai 201210, Peoples R China
关键词
Alanine racemase; Pseudomonas aeruginosa PAO1; Biosynthetic; Homodimer; D-AMINO ACIDS; BACILLUS-STEAROTHERMOPHILUS; ESCHERICHIA-COLI; X-RAY; GROWTH; TUBERCULOSIS; MECHANISM; MUTANTS; BINDS; GENE;
D O I
10.1016/j.bbrc.2018.06.155
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
Alanine racemase is a pyridoxal-5'-phosphate (PLP)-dependent enzyme that reversibly catalyzes the conversion of L-alanine to D-alanine. D-alanine is an essential constituent in many prokaryotic cell structures. Inhibition of alanine racemase is lethal to prokaryotes, creating an attractive target for designing antibacterial drugs. Here we report the crystal structure of biosynthetic alanine racemase (Alr) from a pathogenic bacteria Pseudomonas aeruginosa PAO1. Structural studies showed that P. aeruginosa Alr (PaAlr) adopts a conserved homodimer structure. A guest substrate D-lysine was observed in the active site and refined to dual-conformation. Two buffer ions, malonate and acetate, were bound in the proximity to D-lysine. Biochemical characterization revealed the optimal reaction conditions for PaAlr. (C) 2018 Elsevier Inc. All rights reserved.
引用
收藏
页码:2319 / 2325
页数:7
相关论文
共 40 条
  • [11] The alanine racemase gene is essential for growth of Lactobacillus plantarum
    Hols, P
    Defrenne, C
    Ferain, T
    Derzelle, S
    Delplace, B
    Delcour, J
    [J]. JOURNAL OF BACTERIOLOGY, 1997, 179 (11) : 3804 - 3807
  • [12] Correlation between catalytic activity and monomer-dimer equilibrium of bacterial alanine racemases
    Ju, Jiansong
    Xu, Shujing
    Furukawa, Yukio
    Zhang, Yong
    Misono, Haruo
    Minamino, Tohru
    Namba, Keiichi
    Zhao, Baohua
    Ohnishi, Kouhei
    [J]. JOURNAL OF BIOCHEMISTRY, 2011, 149 (01) : 83 - 89
  • [13] Characterization of endogenous pyridoxal 5′-phosphate-dependent alanine racemase from Bacillus pseudofirmus OF4
    Ju, Jiansong
    Xu, Shujing
    Wen, Jianxin
    Li, Gang
    Ohnishi, Kouhei
    Xue, Yanfen
    Ma, Yanhe
    [J]. JOURNAL OF BIOSCIENCE AND BIOENGINEERING, 2009, 107 (03) : 225 - 229
  • [14] MEGA X: Molecular Evolutionary Genetics Analysis across Computing Platforms
    Kumar, Sudhir
    Stecher, Glen
    Li, Michael
    Knyaz, Christina
    Tamura, Koichiro
    [J]. MOLECULAR BIOLOGY AND EVOLUTION, 2018, 35 (06) : 1547 - 1549
  • [15] D-Amino Acids Govern Stationary Phase Cell Wall Remodeling in Bacteria
    Lam, Hubert
    Oh, Dong-Chan
    Cava, Felipe
    Takacs, Constantin N.
    Clardy, Jon
    de Pedro, Miguel A.
    Waldor, Matthew K.
    [J]. SCIENCE, 2009, 325 (5947) : 1552 - 1555
  • [16] Crystal structure at 1.45 angstrom resolution of alanine racemase from a pathogenic bacterium, Pseudomonas aeruginosa, contains both internal and external aldimine forms
    LeMagueres, P
    Im, H
    Dvorak, A
    Strych, U
    Benedik, M
    Krause, KL
    [J]. BIOCHEMISTRY, 2003, 42 (50) : 14752 - 14761
  • [17] Function of alanine racemase in the physiological activity and cariogenicity of Streptococcus mutans
    Liu, Shiyu
    Wei, Yuan
    Zhou, Xuedong
    Zhang, Keke
    Peng, Xian
    Ren, Biao
    Chen, Vivian
    Cheng, Lei
    Li, Mingyun
    [J]. SCIENTIFIC REPORTS, 2018, 8
  • [18] ORGANIZATION AND EXPRESSION OF THE ESCHERICHIA-COLI K-12 DAD OPERON ENCODING THE SMALLER SUBUNIT OF D-AMINO-ACID DEHYDROGENASE AND THE CATABOLIC ALANINE RACEMASE
    LOBOCKA, M
    HENNIG, J
    WILD, J
    KLOPOTOWSKI, T
    [J]. JOURNAL OF BACTERIOLOGY, 1994, 176 (05) : 1500 - 1510
  • [19] The Versatile Mutational Resistome of Pseudomonas aeruginosa
    Lopez-Causape, Carla
    Cabot, Gabriel
    del Barrio-Tofino, Ester
    Oliver, Antonio
    [J]. FRONTIERS IN MICROBIOLOGY, 2018, 9
  • [20] Phaser crystallographic software
    McCoy, Airlie J.
    Grosse-Kunstleve, Ralf W.
    Adams, Paul D.
    Winn, Martyn D.
    Storoni, Laurent C.
    Read, Randy J.
    [J]. JOURNAL OF APPLIED CRYSTALLOGRAPHY, 2007, 40 : 658 - 674