ATP对羊肉中糖酵解酶磷酸化和乙酰化的影响  

The Role of ATP on Phosphorylation and Acetylation of Glycolytic Enzymes in Lamb Meat

作　　者：董宇 任驰 侯成立[1] 方菲 李欣[1] 张德权[1] DONG Yu;REN Chi;HOU Chengli;FANG Fei;LI Xin;ZHANG Dequan(Institute of Food Science and Technology,Chinese Academy of Agricultural Sciences/Key Laboratory of Agro-products Quality&Safety in Harvest,Storage,Transportation,Management and Control,Ministry of Agriculture and Rural Affairs,Beijing 100193)

机构地区：[1]中国农业科学院农产品加工研究所/农业农村部农产品质量安全收贮运管控重点实验室,北京100193

出　　处：《核农学报》2023年第9期1813-1822,共10页Journal of Nuclear Agricultural Sciences

基　　金：国家自然科学基金重点项目(32030086);中国农业科学院科技创新工程(CAAS-ASTIP-2022-IFST)。

摘　　要：为阐明ATP对肉中糖酵解酶(糖原磷酸化酶、醛缩酶、磷酸丙糖异构酶)磷酸化和乙酰化的影响,以宰后1 h羊背最长肌为试验材料制备肌肉匀浆液,分别添加0、20、50、100 nmol·mg^(-1)蛋白的ATP,构建对照组和低、中、高ATP含量组,在4℃条件下孵育0、1、3、6、12和24 h。采用SDS电泳、蛋白质免疫沉淀、免疫印迹等技术分析ATP和乳酸含量,糖原磷酸化酶、醛缩酶、磷酸丙糖异构酶的磷酸化、乙酰化水平及酶活性变化。结果表明,不同ATP含量处理组的ATP含量显著高于对照组(P<0.05),低ATP含量组的乳酸含量在孵育0~3 h显著低于对照组(P<0.05)。糖原磷酸化酶乙酰化水平、醛缩酶和磷酸丙糖异构酶磷酸化水平分别在孵育0~3、3和0~1 h显著高于对照组(P<0.05)。低、中、高ATP含量组糖原磷酸化酶活性分别在孵育0~1、0~3和0~6 h显著高于对照组(P<0.05)。不同ATP含量组醛缩酶活性在孵育1~6 h显著高于对照组(P<0.05)。低、中、高ATP含量对磷酸丙糖异构酶活性的促进作用分别在1、3、6 h达到最高。综上,添加ATP可在孵育早期通过调控磷酸化和乙酰化修饰促进糖酵解酶活性的提高。本研究结果为宰后早期肌肉中糖酵解酶翻译后修饰及能量代谢相关研究提供了一定的理论基础。In order to elucidate the effect of ATP on phosphorylation and acetylation of glycolytic enzymes(glycogen phosphorylase,aldolase,and triose phosphate isomerase)in meat,muscle homogenate was prepared using sheep longissimus dorsi muscle at 1 h postmortem.The muscle homogenate was incubated with 0,20,50,and 100 nmol·mg^(-1) protein ATP to obtain control,low,medium,and high ATP content groups,respectively.The samples were incubated at 4℃for 0,1,3,6,12 and 24 h.The phosphorylation levels,acetylation levels and enzyme activity of glycogen phosphorylase,aldolase,and triose phosphate isomerase as well as the contents of ATP and lactate were measured by SDS electrophoresis,protein immunoprecipitation and Western blotting.The results showed that the ATP contents in the different ATP content treatment groups were significantly higher than those in the control group(P<0.05),and the lactate contents of the low ATP content group were significantly lower than those in the control group at 0-3 h of incubation(P<0.05).The acetylation levels of glycogen phosphorylase,the phosphorylation levels of aldolase and triose phosphate isomerase at 0-3,3 and 0-1 h of incubation were significantly higher than those in the control group(P<0.05).The activities of glycogen phosphorylase in the low,medium and high ATP content groups were significantly higher than that in the control group within 0-1,0-3 and 0-6 h of incubation respectively(P<0.05).The activities of aldolase of the different ATP content group were significantly higher than those in the control group at 1-6 h of incubation(P<0.05).The promoting effect of low,medium and high ATP content on triose phosphate isomerase activity reached the highest at 1,3 and 6 h,respectively.In conclusion,the addition of ATP can promote glycolytic enzyme activity by regulating phosphorylation and acetylation levels in the early incubation period.The results of this study provide a theoretical basis for carring out research about glycolytic enzyme post-translational modifications and energy metabol

关 键 词：肉 ATP 糖酵解酶 磷酸化 乙酰化 

分 类 号：TS251.53[轻工技术与工程—农产品加工及贮藏工程]