Amorphin is phosphorylase; Phosphorylase is an alpha-actinin-binding protein

被引:36
作者
Chowrashi, P [1 ]
Mittal, B [1 ]
Sanger, JM [1 ]
Sanger, JW [1 ]
机构
[1] Univ Penn, Sch Med, Dept Cell & Dev Biol, Philadelphia, PA 19104 USA
来源
CELL MOTILITY AND THE CYTOSKELETON | 2002年 / 53卷 / 02期
关键词
muscle; sarcomere; Z-band; enzyme; thin filaments; McArdle's Disease; Green Fluorescent Protein;
D O I
10.1002/cm.10059
中图分类号
Q2 [细胞生物学];
学科分类号
071009 ; 090102 ;
摘要
In a study of myofibrillar proteins, Chowrashi and Pepe [1982: J. Cell Biol. 94:565-573] reported the isolation of a new, 85-kD Z-band protein that they named amorphin. We report that partial sequences of purified amorphin protein indicate that amorphin is identical to phosphorylase, an enzyme important in the metabolism of glycogen. Anti-amorphin antibodies also reacted with purified chicken and rabbit phosphorylase. To explore the basis for phosphorylase's (amorphin's) localization in the Z-bands of skeletal muscles, we reacted biotinylated alpha-actinin with purified amorphin and with purified phosphorylase and found that alpha-actinin bound to each. Radioimmune assays also indicated that phosphorylase (amorphin) bound to alpha-actinin, and, with lower affinity, to F-actin. Negative staining of actin filaments demonstrated that alpha-actinin mediates the binding of phosphorylase to actin filaments. There are several glycolytic enzymes that bind actin (e.g., aldolase, phosphofructokinase, and pyruvate kinase), but phosphorylase is the first one demonstrated to bind alpha-actinin. Localization of phosphorylase in live cells was assessed by transfecting cultures of quail embryonic myotubes with plasmids expressing phosphorylase fused to Green Fluorescent Protein (GFP). This resulted in targeting of the fusion protein to Z-bands accompanied by a diffuse pattern in the cytoplasm. Cell Motil. Cytoskeleton 53: 125-135, 2002. (C) 2002 Wiley-Liss, Inc.
引用
收藏
页码:125 / 135
页数:11
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