CAJ | 학술논문

钙蛋白酶抑制蛋白(Calpastatin,CAST)是一种内源性的需要Ca~(2+)激活的钙蛋白酶抑制剂,在肌肉组织的蛋白质降解过程中起重要的调节作用,文章利用牛CAST基因的mRNA序列,通过逆转录RT-PCR首次克隆获得绵羊CAST基因2型转录本和4型转录本的部分cDNA序列,并对序列进行生物信息学分析.CAST基因2型转录本的扩增片段为4385 bp,完整的开放阅读框为2 361 bp,编码786个氨基酸;CAST基因4型转录本的扩增片段为1 467 bp,完整的开放阅读框为1 317 bp,编码438个氨基酸.CASTⅡ型蛋白序列存在4个保守结构域,CASTⅣ型蛋白序列存在3个保守结构域;两者的二级结构均以螺旋为主,富含疏水区域,其氨基酸序列存在多个磷酸化位点以及蛋白激酶C(Protein kinase C,PKC)的磷酸化位点.通过RT-PCR分析CAST基因2型转录本和4型转录本的组织表达谱,结果表明CAST基因2型转录本在所检测的10个组织中均表达,CAST基因4型转录本仅在睾丸组织中表达.
개단백매억제단백(Calpastatin,CAST)시일충내원성적수요Ca~(2+)격활적개단백매억제제,재기육조직적단백질강해과정중기중요적조절작용,문장이용우CAST기인적mRNA서렬,통과역전록RT-PCR수차극륭획득면양CAST기인2형전록본화4형전록본적부분cDNA서렬,병대서렬진행생물신식학분석.CAST기인2형전록본적확증편단위4385 bp,완정적개방열독광위2 361 bp,편마786개안기산;CAST기인4형전록본적확증편단위1 467 bp,완정적개방열독광위1 317 bp,편마438개안기산.CASTⅡ형단백서렬존재4개보수결구역,CASTⅣ형단백서렬존재3개보수결구역;량자적이급결구균이라선위주,부함소수구역,기안기산서렬존재다개린산화위점이급단백격매C(Protein kinase C,PKC)적린산화위점.통과RT-PCR분석CAST기인2형전록본화4형전록본적조직표체보,결과표명CAST기인2형전록본재소검측적10개조직중균표체,CAST기인4형전록본부재고환조직중표체.
As an endogenous inhibitor of the calpain system activated by Ca~(2+), calpastatin (CAST) plays a regulatory role in muscle proteolysis. Based on the bovine mRNA sequences, part of cDNA fragments of sheep CAST transcript 2 and 4 were obtained by RT-PCR. Bioinformatic analysis showed that sheep CAST transcript 2 was 4 358 bp in length with an open reading frame (ORF) 2 361 bp long and encoded 786 amino acids, while sheep CAST transcript 4 was 1 467 bp in length with 1 317 bp ORF encoding 438 amino acids. It was predicted thatCAST type II contained four conserved domains and CAST type IV contained three conserved domains, and their secondary structures were rich in both hydrophobic regions and helical regions, with certain conserved phosphorylation sites and phosphorylation sites of protein kinase C (PKC). RT-PCR was conducted to analyze the expression patterns of CAST transcript 2 and transcript 4. CAST transcript 2 was expressed in ten tissues detected while CAST transcript 4 only in testis.