論文 - 詳細
RRC ID | 30573 |
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著者 | Yamamoto K, Usuda K, Kakuta Y, Kimura M, Higashiura A, Nakagawa A, Aso Y, Suzuki M. |
タイトル | Structural basis for catalytic activity of a silkworm Delta-class glutathione transferase. |
ジャーナル | Biochim Biophys Acta |
Abstract |
BACKGROUND:Glutathione transferase (GST) catalyzes glutathione conjugation, a major detoxification pathway for xenobiotics and endogenous substances. Here, we determined the crystal structure of a Delta-class GST from Bombyx mori (bmGSTD) to examine its catalytic residues. METHODS:The three-dimensional structure of bmGSTD was resolved by the molecular replacement method and refined to a resolution of 2.0Å. RESULTS:Structural alignment with a Delta-class GST of Anopheles gambiae indicated that bmGSTD contains 2 distinct domains (an N-terminal domain and a C-terminal domain) connected by a linker. The bound glutathione localized at the N-terminal domain. Putative catalytic residues were changed to alanine by site-directed mutagenesis, and the resulting mutants were characterized in terms of catalytic activity using glutathione and 1-chloro-2,4-dinitrobenzene, a synthetic substrate of GST. Kinetic analysis of bmGSTD mutants indicated that Ser11, Gln51, His52, Ser67, and Arg68 are important for enzyme function. GENERAL SIGNIFICANCE:These results provide structural insights into the catalysis of glutathione conjugation in B. mori by bmGSTD. |
巻・号 | 1820(10) |
ページ | 1469-74 |
公開日 | 2012-10-1 |
DOI | 10.1016/j.bbagen.2012.04.022 |
PII | S0304-4165(12)00127-4 |
PMID | 22579926 |
MeSH | Amino Acid Sequence Animals Bombyx / enzymology* Bombyx / genetics Catalysis Catalytic Domain / genetics Crystallization Enzyme Activation Enzyme Stability / genetics Glutathione Transferase / chemistry* Glutathione Transferase / classification Glutathione Transferase / genetics Glutathione Transferase / metabolism* Models, Molecular Molecular Sequence Data Mutagenesis, Site-Directed Protein Interaction Domains and Motifs / genetics Sequence Homology, Amino Acid Structure-Activity Relationship |
IF | 3.411 |
引用数 | 16 |
WOS 分野 | BIOPHYSICS BIOCHEMISTRY & MOLECULAR BIOLOGY |
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