First structure of archaeal branched-chain amino acid aminotransferase from Thermoproteus uzoniensis specific for L-amino acids and R-amines

被引：24

作者：

Boyko, Konstantin M. ^{[1
,2
]}

Stekhanova, Tatiana N. ^{[1
]}

Nikolaeva, Alena Yu ^{[1
,2
]}

Mardanov, Andrey V. ^{[3
]}

Rakitin, Andrey L. ^{[3
]}

Ravin, Nikolai V. ^{[3
]}

Bezsudnova, Ekaterina Yu ^{[1
]}

Popov, Vladimir O. ^{[1
,2
]}

机构：

[1] Russian Acad Sci, Biotechnol Res Ctr, AN Bach Inst Biochem, Leninsky Ave 33,Bld 2, Moscow 119071, Russia

[2] Natl Res Ctr Kurchatov Inst, NBICS Ctr, Akad Kurchatova Sqr 1, Moscow 123182, Russia

[3] Russian Acad Sci, Biotechnol Res Ctr, Inst Bioengn, Leninsky Ave 33,Bld 2, Moscow 119071, Russia

来源：

EXTREMOPHILES | 2016年 / 20卷 / 02期

基金：

俄罗斯科学基金会;

关键词：

Archaea; Branched-chain amino acid aminotransferase; Selectivity; 3D structure; Thermostability; SUBSTRATE RECOGNITION; CRYSTAL-STRUCTURES; COMPLEXES; GLUTAMATE; SEQUENCE; ENZYME; TOOL;

D O I：

10.1007/s00792-016-0816-z

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：

The gene TUZN1299 from the genome of the hyperthermophilic archaeon Thermoproteus uzoniensis encoding a new 32.8 kDa branched-chain amino acid aminotransferase (BCAT) was expressed in Escherichia coli. The recombinant protein TUZN1299 was purified to homogeneity in the PLP-bound form. TUZN1299 was active towards branched-chain amino acids (L-Val, L-Leu, L-Ile) and showed low but detectable activity toward (R)-alpha-methylbenzylamine. The enzyme exhibits hightemperature optimum, thermal stability, and tolerance to organic solvents. The structure of an archaeal BCAT called TUZN1299 was solved for the first time (at 2.0 angstrom resolution). TUZN1299 has a typical BCAT type IV fold, and the organization of its active site is similar to that of bacterial BCATs. However, there are some differences in the amino acid composition of the active site.

引用

页码：215 / 225

页数：11

共 35 条

[1]

[Anonymous], PROTEIN ADAPTATION I

[2]

Bateman A, 2002, NUCLEIC ACIDS RES, V30, P276, DOI [10.1093/nar/gkp985, 10.1093/nar/gkh121, 10.1093/nar/gkr1065]

[3] The Discovery of in Vivo Active Mitochondrial Branched-Chain Aminotransferase (BCATm) Inhibitors by Hybridizing Fragment and HTS Hits [J].

Bertrand, Sophie M. ;

Ancellin, Nicolas ;

Beaufils, Benjamin ;

Bingham, Ryan P. ;

Borthwick, Jennifer A. ;

Boullay, Anne-Benedicte ;

Boursier, Eric ;

Carter, Paul S. ;

Chung, Chun-wa ;

Churcher, Ian ;

Dodic, Nerina ;

Fouchet, Marie-Helene ;

Foumier, Charlene ;

Francis, Peter L. ;

Gummer, Laura A. ;

Herry, Kenny ;

Hobbs, Andrew ;

Hobbs, Clare I. ;

Homes, Paul ;

Jamieson, Craig ;

Nicodeme, Edwige ;

Pickettlt, Stephen D. ;

Reid, Fain H. ;

Simpson, Graham L. ;

Sloan, Lisa A. ;

Smith, Sarah E. ;

Somers, Donald O'N. ;

Spitzfaden, Claus ;

Suckling, Cohn J. ;

Valko, Kara ;

Washio, Yoshiaki ;

Young, Robert J. .

JOURNAL OF MEDICINAL CHEMISTRY, 2015, 58 (18) :7140-7163

[4] From gene to structure: The protein factory of the NBICS Centre of Kurchatov Institute [J].

Boyko, K. M. ;

Lipkin, A. V. ;

Popov, V. O. ;

Kovalchuk, M. V. .

CRYSTALLOGRAPHY REPORTS, 2013, 58 (03) :442-449

[5] Structural analysis of mycobacterial branched-chain aminotransferase: implications for inhibitor design [J].

Castell, Alina ;

Mille, Christian ;

Unge, Torsten .

ACTA CRYSTALLOGRAPHICA SECTION D-STRUCTURAL BIOLOGY, 2010, 66 :549-557

[6] Crystal Structures of Complexes of the Branched-Chain Aminotransferase from Deinococcus radiodurans with α - Ketoisocaproate and L-Glutamate Suggest the Radiation Resistance of This Enzyme for Catalysis [J].

Chen, Chung-De ;

Lin, Chih-Hao ;

Chuankhayan, Phimonphan ;

Huang, Yen-Chieh ;

Hsieh, Yin-Cheng ;

Huang, Tien-Feng ;

Guan, Hong-Hsiang ;

Liu, Ming-Yih ;

Chang, Wen-Chang ;

Chen, Chun-Jung .

JOURNAL OF BACTERIOLOGY, 2012, 194 (22) :6206-6216

[7]

Collaborative Computational Project, 1994, ACTA CRYSTALLOGR D, V50, P760, DOI DOI 10.1107/S0907444994003112

[8] Improved R-factors for diffraction data analysis in macromolecular crystallography [J].

Diederichs, K ;

Karplus, PA .

NATURE STRUCTURAL BIOLOGY, 1997, 4 (04) :269-275

[9] Features and development of Coot [J].

Emsley, P. ;

Lohkamp, B. ;

Scott, W. G. ;

Cowtan, K. .

ACTA CRYSTALLOGRAPHICA SECTION D-BIOLOGICAL CRYSTALLOGRAPHY, 2010, 66 :486-501

[10] Scaling and assessment of data quality [J].

Evans, P .

ACTA CRYSTALLOGRAPHICA SECTION D-STRUCTURAL BIOLOGY, 2006, 62 :72-82

← 1 2 3 4 →