The Role of Amino Acid Residues in the Active Site of L-Methionine γ-lyase from Pseudomonas putida

• FUKUMOTO Mitsuki

  Graduate School of Natural Science and Technology, Okayama University Graduate School of Natural Science and Technology, Okayama University

• KUDOU Daizou

  Graduate School of Natural Science and Technology, Okayama University Graduate School of Natural Science and Technology, Okayama University

• MURANO Shouko

  Graduate School of Science and Technology, Department of Applied Biology, Kyoto Institute of Technology Graduate School of Science and Technology, Department of Applied Biology, Kyoto Institute of Technology

• SHIBA Tomoo

  Graduate School of Science and Technology, Department of Applied Biology, Kyoto Institute of Technology Graduate School of Science and Technology, Department of Applied Biology, Kyoto Institute of Technology

• SATO Dan

  Institute for Advanced Biosciences, Keio University Institute for Advanced Biosciences, Keio University

• TAMURA Takashi

  Graduate School of Natural Science and Technology, Okayama University Graduate School of Natural Science and Technology, Okayama University

• HARADA Shigeharu

  Graduate School of Science and Technology, Department of Applied Biology, Kyoto Institute of Technology Graduate School of Science and Technology, Department of Applied Biology, Kyoto Institute of Technology

• INAGAKI Kenji

  Graduate School of Natural Science and Technology, Okayama University Graduate School of Natural Science and Technology, Okayama University

書誌事項

タイトル別名

• The Role of Amino Acid Residues in the Active Site of <small>L</small>-Methionine γ-lyase from <i>Pseudomonas putida</i>
• The Role of Amino Acid Residues in the Active Site of<scp>L</scp>-Methionine γ-lyase from<i>Pseudomonas putida</i>
• The role of amino acid residues in the active site of l-methionine gamma-lyase from Pseudomonas putida
• The role of amino acids residues in the active site of l-methionine γ-lyase from Pseudomonas putida

抄録

Cys116, Lys240^*, and Asp241^* (asterisks indicate residues from the second subunit of the active dimer) at the active site of L-methionine γ-lyase of Pseudomonas putida (MGL_Pp) are highly conserved among heterologous MGLs. In a previous study, we found that substitution of Cys116 for His led to a drastic increase in activity toward L-cysteine and a decrease in that toward L-methionine. In this study, we examined some properties of the C116H mutant by kinetic analysis and 3D structural analysis. We assumed that substitution of Cys116 for His broke the original hydrogen-bond network and that this induced a significant effect of Tyr114 as a general acid catalyst, possibly due to the narrow space in the active site. The C116H mutant acquired a novel β-elimination activity and lead a drastic conformation change in the histidine residue at position 116 by binding the substrate, suggesting that this His residue affects the reaction specificity of C116H. Furthermore, we suggest that Lys240^* is important for substrate recognition and structural stability and that Asp241^* is also involved in substrate specificity in the elimination reaction. Based on this, we suggest that the hydrogen-bond network among Cys116, Lys240^*, and Asp241^* contributes to substrate specificity that is, to L-methionine recognition at the active site in MGL_Pp.

収録刊行物

• Ｂｉｏｓｃｉｅｎｃｅ，　Ｂｉｏｔｅｃｈｎｏｌｏｇｙ，　ａｎｄ　Ｂｉｏｃｈｅｍｉｓｔｒｙ

  Ｂｉｏｓｃｉｅｎｃｅ，　Ｂｉｏｔｅｃｈｎｏｌｏｇｙ，　ａｎｄ　Ｂｉｏｃｈｅｍｉｓｔｒｙ 76 (7), 1275-1284, 2012

  公益社団法人 日本農芸化学会

キーワード

• <small>L</small>-methionine γ-lyase
• crystal structure
• mutagenesis
• reaction specificity

詳細情報

• CRID: 1390282681455281920
• NII論文ID: 130001862713
• NII書誌ID: AA10824164
• DOI: 10.1271/bbb.110906
• COI: 1:STN:280:DC%2BC38jpt1KjtA%3D%3D
• ISSN: 13476947
• ISSN: 09168451
• NDL書誌ID: 023840605
• PubMed: 22785484
• Web Site: https://ndlsearch.ndl.go.jp/books/R000000004-I023840605
• Web Site: http://www.tandfonline.com/doi/pdf/10.1271/bbb.110906
• 本文言語コード: en
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• 抄録ライセンスフラグ: 使用不可