Affinity labeling of muscle phosphorylase b with .ALPHA.-cyclodextrin-dialdehyde.

• TAKAGI Shiro

  Department of Agricultural Chemistry, Faculty of Agriculture, Tohoku University

• KOBAYASHI Mikihiko

  Department of Agricultural Chemistry, Faculty of Agriculture, Tohoku University

• URAYAMA Tadanori

  Department of Agricultural Chemistry, Faculty of Agriculture, Tohoku University

• SUZAWA Itsuko

  Department of Agricultural Chemistry, Faculty of Agriculture, Tohoku University

• MATSUDA Kazuo

  Department of Agricultural Chemistry, Faculty of Agriculture, Tohoku University Present address: Iwaki-Meisei University

• ICHISHIMA Eiji

  Department of Agricultural Chemistry, Faculty of Agriculture, Tohoku University

抄録

Rabbit muscle phosphorylase b was modified with a substrate analog, the 2', 3'-dialdehyde derivative of α-cyclodextrin (dial-α-CD). Although the inhibition of phosphorylase b by α-, β-, and γ-cyclodextrins gave rather high Ki values (10-25 ITIM), the dial-CD gave much smaller Ki values of 1.2-3.5mM. Moreover, the latter inhibition was time-dependent and accelerated by higher pHs and higher concentrations of dial-CD. Incorporation of the dial-CD into the enzyme was proportional to the loss of enzyme activity and became stationary at about 1 mol of dial-CD bound to a mol of enzyme subunit. Modification was greatly suppressed by the presence of substrate glycogen. Glucose 1-phosphate was not effective. The dial-CD-modified phosphorylase b was purified by Sephadex G-75 and Con A-Sepharose column chromatography. The modified enzyme gave a single band of activity having a K_app of 6.3% glycogen on affinity gel electrophoresis, which showed that the modified enzyme had a very low affinity for glycogen. Comparison of Km values of the native and modified phosphorylase b showed that the Km values for the glucan substrates increased 8 to 11-fold in the modified enzyme. These results suggested that the glycogen storage site of muscle phosphorylase b might be modified with dial-CD and the modified enzyme significantly decreased in the affinity for the substrate glycogen.

収録刊行物

• Agricultural and Biological Chemistry

  Agricultural and Biological Chemistry 52 (11), 2709-2716, 1988

  公益社団法人 日本農芸化学会

キーワード

• General Agricultural and Biological Sciences
• General Biochemistry, Genetics and Molecular Biology

詳細情報

• CRID: 1390001206464538112
• NII論文ID: 110006323529
• NII書誌ID: AA00515312
• DOI: 10.1271/bbb1961.52.2709
• COI: 1:CAS:528:DyaL1MXntFKrtg%3D%3D
• ISSN: 18811280
• ISSN: 00021369
• 本文言語コード: en
• データソース種別:
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• 抄録ライセンスフラグ: 使用不可