Role of the Pyrrolidine Ring of Proline in Determining the Substrate Specificity of cdc2 Kinase or cdk5.
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概要
- 論文の詳細を見る
To examine structural features of proline which are essential for the proline-directed phosphorylation by cdc2 kinase or cdk5, we prepared the peptide representing the cdc2 kinase phosphorylation site at Ser-55 in vimentin [Ser-Leu-Tyr-Ser-Ser-Ser55-Pro56-Gly-Gly58-Ala-Tyr-NH2], the peptide containing arginine in place of Gly-58, and their derivatives containing various N-methylamino acids or proline homologs in place of Pro-56, and tested them as substrates for the kinases. While substitution of the proline by proline homologs (L-pipecolic acid or L-azetidine-2-carboxylic acid) increased the Km value 2- to 4-fold at utmost, substitution by N-methylamino acids (sarcosine, L-N-methylalanine, L-N-methylvaline, or L-N-methylleucine) increased the Km value 7- to 40-fold for cdc2 kinase. For cdk5, these substitutions led to parallel effects on the Km value to those found for cdc2 kinase; cdk5 recognized the peptides with a proline specificity similar to that for cdc2 kinase. These results suggest that the pyrrolidine ring of proline is important for substrate recognition by cdc2 kinase or cdk5. Molecular dynamics and molecular mechanics simulations indicated that the pyrrolidine ring of proline is optimal to stabilize a β-turn at the phosphorylation site and that the Km values of the peptides for the enzymes might be related to the probability of the turn structure. The results obtained here also suggest that the pyrrolidine ring of proline is required to maintain a high Vmax value for cdc2 kinase or especially for cdk5. These will aid in designing specific substrates or inhibitors for cdc2 kinase or cdk5.
- 社団法人 日本生化学会の論文
著者
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Inagaki Masaki
Laboratory Of Biochemistry Aichi Cancer Center Research Institute
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Hisanaga Shin-ichi
Laboratory Of Cell And Developmental Biology Faculty Of Bioscience Tokyo Institute Of Technology
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Ando Shoji
Laboratory Of Chemistry Saga Medical School
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Sasaki Yasuharu
Life Science Research Center Asahi Chemical Industry Co. Ltd.
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ITO Hikaru
Laboratory of Biochemistry, Aichi Cancer Center Research Institute
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Kamata Tomoko
Analytical Research And Computer Science Center Asahi Chemical Industry Co. Ltd.
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Kishimoto Takeo
Laboratory Of Cell And Developmental Biology Department Of Biological Information Graduate School Of
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Ikuhara Toshihiko
Life Science Research Center Asahi Chemical Industry Co. Ltd.
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