軟体動物平滑筋Ca 結合蛋白質の単離とアミノ酸配列解析

概要

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原著To elucidate the intracellular localization of a Ca-binding protein，MCBP‐450，andits regulatory mechanism of contraction，it has been thought that an experiment using immunoelectron microscopy would be valid and significant．As the first step，in the present study，the isolation and purification of MCBP‐450 from the anterior byssus retractor muscle （ABRM）of Mytilus edulis were examined．Based on the method reported by Yamanobe and Sugi （1993）1），a sample containing a protein with a molecular weight corresponding to 450 kDa was separated by SDS-PAGE．It was proved that the protein had the ability to bind Ca ions by detecting the fluorescence emission of Ca-indicator quin2．It is reasonable to conclude that the sample contains MCBP-450． SDS‐PAGE after the dialysis of purified samples showed no distinct band corresponding to 450 kDa，but a marked band corresponding to ～100 kDa. The amino acid sequence of three poly‐peptides fragmented enzymatically from the ～100 kDa protein was examined with the Procise 494HT Protein Sequencing System，and the sequences of 12 amino acids were determined respectively．A homology search of these sequences for the various known protein sequences using the NCBInr Library indicated high homology（～90％）with α‐actinin in various kinds of cells and tissues．It was suggested that part of the MCBP-450 molecule resembles closely some parts of α‐actinin．
2012-06-30