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鸡隐钙素同源物的鉴定与发育表达

Identification and developmental expression of a chicken calsequestrin homolog.

作者信息

Choi E S, Clegg D O

机构信息

Department of Biological Sciences, University of California, Santa Barbara 93106.

出版信息

Dev Biol. 1990 Nov;142(1):169-77. doi: 10.1016/0012-1606(90)90160-k.

Abstract

Calsequestrin (CAL) is a calcium-binding protein whose primary function is thought to involve sequestration of calcium in the muscle sarcoplasmic reticulum (SR). Little is known about the mechanisms regulating CAL expression, or about the role of this protein in muscle development. In addition, CAL may regulate calcium localization in some nonmuscle cells. We have identified an avian calsequestrin homolog. The predicted amino acid sequence of the avian CAL, first described as a laminin binding protein, and named aspartactin, is 70-80% identical to mammalian CAL sequences. We have used affinity-purified antibodies and cDNA probes to investigate expression in developing and adult chicken tissues. In adult chickens, the avian CAL homolog was expressed in slow and fast twitch skeletal muscle as well as in cardiac muscle. Surprisingly high levels of CAL protein were also detected in cerebellum. During development, CAL mRNA and protein were detected in Embryonic Day 5 (E-5) limb primordia, well before the initiation of myoblast fusion. In leg skeletal muscle, CAL protein and mRNA increase approximately 10-fold from E-8 to E-18 with a time course that just precedes myoblast fusion. This early expression pattern was also observed in cultured chicken pectoral myoblasts, and appears to be regulated at the level of mRNA abundance. The developmental profile of CAL expression is compared to that of other muscle proteins and possible additional functions of CAL are discussed.

摘要

肌集钙蛋白(CAL)是一种钙结合蛋白,其主要功能被认为是将钙隔离在肌肉肌浆网(SR)中。关于调节CAL表达的机制,或者这种蛋白质在肌肉发育中的作用,人们所知甚少。此外,CAL可能在一些非肌肉细胞中调节钙的定位。我们已经鉴定出一种禽类肌集钙蛋白同源物。禽类CAL的预测氨基酸序列最初被描述为一种层粘连蛋白结合蛋白,并命名为天冬氨酸肌动蛋白,与哺乳动物CAL序列有70%-80%的同一性。我们使用亲和纯化抗体和cDNA探针来研究其在发育中和成年鸡组织中的表达。在成年鸡中,禽类CAL同源物在慢肌和快肌骨骼肌以及心肌中表达。令人惊讶的是,在小脑中也检测到了高水平的CAL蛋白。在发育过程中,在胚胎第5天(E-5)的肢体原基中检测到CAL mRNA和蛋白,早于成肌细胞融合的开始。在腿部骨骼肌中,CAL蛋白和mRNA从E-8到E-18增加约10倍,其时间进程刚好先于成肌细胞融合。这种早期表达模式在培养的鸡胸肌成肌细胞中也观察到,并且似乎在mRNA丰度水平上受到调节。将CAL表达的发育情况与其他肌肉蛋白的发育情况进行了比较,并讨论了CAL可能的其他功能。

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