摘要
以6月龄滩羊背最长肌为研究对象,测定4℃贮藏0,96,192 h时剪切力、线粒体Ca^(2+)水平、钙蛋白酶-1及细胞凋亡效应酶-3活性。基于同位素标记相对和绝对定量技术(iTRAQ)结合多维液相色谱-串联质谱(LC-MS/MS)技术,研究贮藏期间Ca^(2+)信号通路及凋亡调节信号转导通路蛋白质表达量的变化情况。结果表明:4℃贮藏期间,滩羊肉剪切力先降低后升高,线粒体Ca^(2+)含量先升高后降低,钙蛋白酶-1活性持续降低,细胞凋亡效应酶-3活性先上升后下降。滩羊肉剪切力与Ca^(2+)水平呈显著负相关,与钙蛋白酶-1活性呈极显著正相关,与细胞凋亡效应酶-3活性呈极显著负相关。蛋白质组学分析表明:Ca^(2+)跨膜转运ATP酶利用ATP水解的能量驱动Ca^(2+)跨膜运输,电压依赖性阴离子通道蛋白贮藏初期上调表达,为Ca^(2+)流入线粒体提供渠道,导致线粒体Ca^(2+)超载、线粒体功能紊乱,使线粒体复合体表达量上升。同时,VDAC异常表达促使CYTC释放至胞浆,最终激活细胞凋亡效应酶-3,水解结构蛋白质。另外,Ca^(2+)可间接导致肌动蛋白表达量下调,引起细胞收缩,从而影响肉的嫩度。本研究结果证实Ca^(2+)在肉品嫩度形成过程中起重要的信号转导作用。
This study was design to clarify the important transduction role of Ca^(2+)in meat tenderness development of Tan sheep during postmortem storage.Results indicated that the shear force decreased significantly before 96 h,and then increased significantly from 96-192 h.Mitochondrial Ca^(2+)levels increased significantly before 96 h,and decreased significantly after 96 h.The activity of calpain-1 decreased significantly with increasing time postmortem.The activity of caspase-3 increased significantly before 96 h and then decreased significantly.Correlation analysis indicated that the shear force was positively correlated with mitochondrial Ca^(2+)levels,and significant positively correlated with activity of caspase-3,significant negatively correlated with activity of calpain-1.0-96 h,5 up-regulated differentially abundant proteins and 2 down-regulated differentially abundant proteins were enriched in Ca^(2+)signaling pathway.96-192 h,2 down-regulated differentially abundant proteins were enriched in Ca^(2+)signaling pathway.0-96 h,30 differentially abundant proteins were enriched in apoptosis signal transduction pathways.96-192 h,15 differentially abundant proteins were enriched in apoptosis signal transduction pathways.The energy from hydrolysis of ATP was used by SERCA to drive Ca^(2+)transport across membranes.Increasing of VDAC likely provided channels for Ca^(2+)lead to mitochondrial Ca^(2+)overloading and mitochondrial dysfunction.Meanwhile,increasing of VDAC could be related to the increase of CYTC in cytoplasm,thereby caused the cascade of apoptosis,and activated caspase-3 to hydrolyze structural proteins,finally influence tenderness of Tan sheep meat.In addition,the decrease of actin was caused by Ca^(2+),and provided the contractile mechanism of muscle tissue,thus affecting the tenderness of meat.The conclusion of this study further confirmed the important signal transduction role of Ca^(2+)in the development of meat tenderness during postmortem storage.
作者
姬琛
尤丽琴
刘吉娟
罗瑞明
Ji Chen;You Liqin;Liu Jijuan;Luo Ruiming(College of Agriculture,Ningxia University,Yinchuan 750021;College of Food and Wine,Ningxia University,Yinchuan 750021)
出处
《中国食品学报》
EI
CAS
CSCD
北大核心
2022年第12期214-223,共10页
Journal of Chinese Institute Of Food Science and Technology
基金
国家自然科学基金地区基金项目(31860427)。
作者简介
第一作者:姬琛,女,博士生;通信作者:罗瑞明,E-mail:11003912921tfboys@163.com。
