蛋白质磷酸化调控羊肉肌原纤维蛋白的功能

doi:10.3864/j.issn.0578-1752.2016.07.012

Abstract

Abstract: 【Objective】The aim of this study was to analyze the effect of the phosphorylation level which was improved by activating the activity of protein kinase on myofibrillar proteins’ degradation and contraction.【Method】Improve the activity of protein kinases by adding protein kinase A activator Forskolin and protein kinase C activator PMA, followed by the increasing of the phosphorylation level of myofirillar proteins in mutton muscle. By comparing the myofibrillar fragmentation index (MFI), the degree of myofibrillar degradation, the sarcomere length, and other indicators between activator groups and control group to determine the effect of protein phosphorylation levels on lamb muscle contraction and degradation. 【Result】The activity of protein kinases in the PMA group (PKC activated group) were higher than the control group after 1 hour, 2 hour, and 4 hour incubation (P＜0.05). The activity of protein kinases in the Forskolin group (PKA activated group) were higher than in the control group after 1 hour and 4 hour incubation (P＜0.05). The highest activity of the protein kinases appeared after 1 hour incubation. PMA and Forskolin can improve the phosphorylation level of titin, myosin binding protein C, tropomyosin, myosin light chain 2, and a protein of 25 kDa by activating the protein kinases, but the phosphorylation level of myosin heavy chain and actin had no changes. The degradation degree of the PMA group and the Forskolin group were lower than the control group, and the sarcomere length of the PMA group and the Forskolin group was shorter than the control group. 【Conclusion】 On the one hand, the improvement of the myofirillar proteins phosphorylation level can reduce the degree of myofibrillar degradation, while on the other hand, the improvement of the myofirillar proteins phosphorylation level may enhance the force of muscle contraction through the effect on myosin light chain 2, may accelerate the contraction of filament through the effect on adjacent tropomyosin, and finally influence the rigor mortis and tenderization.

Key words: mutton, protein phosphorylation, myofibrillar proteins, muscle contraction, protein degradation

CHEN Li-juan, LI Xin, LI Zheng, LI Pei-di, LI Zhong-wen, ZHANG De-quan. Protein Phosphorylation on the Function of Myofibrillar Proteins in Mutton Muscle[J].Scientia Agricultura Sinica, 2016, 49(7): 1360-1370.

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References

[1] 钱小红, 贺福初. 蛋白质组学: 理论与方法. 北京: 北京科学出版社, 2003.

Qian X H, He F C. Proteomics: Theory and Methods. Beijing: Beijing Science Press, 2003. (in Chinese)

[2] Cohen P. The origins of protein phosphorylation. Nature Cell Biology, 2002, 4(5): E127-E130.

[3] Heeley D H, Watson M H, Mak A S, Dubord P, Smillie L B. Effect of phosphorylation on the interaction and functional properties of rabbit striated-muscle alpha-alpha-tropomyosin. Journal of Biological Chemistry, 1989, 264(5): 2424-2430.

[4] Mazzei G J, Kuo J F. Phosphorylation of skeletal-muscle troponin I and troponin T by phospholipid-sensitive Ca²⁺-dependent protein kinase and its inhibition by troponin C and tropomyosin. Biochemical Journal, 1984, 218(2): 361-369.

[5] Ryder J W, Lau K S, Kamm K E, Stull J T. Enhanced skeletal muscle contraction with myosin light chain phosphorylation by a calmodulin-sensing kinase. Journal of Biological Chemistry, 2007, 282(28): 20447-20454.

[6] Zhi G, Ryder J W, Huang J, Ding P, Chen Y, Zhao Y, Kamm K E, Stull J T. Myosin light chain kinase and myosin phosphorylation effect frequency-dependent potentiation of skeletal muscle contraction. Proceedings of the National Academy of Sciences of the United States of America, 2005, 102(48): 17519-17524.

[7] Stull T J, Kamm K E, Vandenboom R. Myosin light chain kinase and the role of myosin light chain phosphorylation in skeletal muscle. Archives of Biochemistry and Biophysics, 2011, 510: 120-128.

[8] Lisa F D, Tullio R D, Salamino F, Barboto R, Melloni E, Siliprandi N, Schiaffino S, Pontremoli S. Specific degradation of troponin T and I by mu-calpain and its modulation by substrate phosphorylation. Biochemical Journal, 1995, 308: 57-61.

[9] Teruhiko Toyo-oka. Phosphorylation with cyclic adenosine 3’: 5’monophosphate-dependent protein kinase renders bovine cardiac troponin sensitive to the degradation by calcium-activated neutral protease. Biochemical and Biophysical Research Communications, 1982, 107(1): 44-50.

[10] Doumit M E, Bates R O. Regulation of pork water holding capacity, color, and tenderness by protein phosphorylation. Pork Quality, 2000.

[11] 张大雷, 刘芳明, 杨树龙, 杨蓓, 邹挺. PKA和PKC系统对小鼠精原细胞增殖的影响. 南昌大学学报: 理科版, 2011, 35(5): 476-479.

Zhang D L, Liu M F, Yang S L, Yang B, Zou T. The effect of PKA and PKC on spermatagonial cells proliferation in mice. Journal of Nanchang University: Natural Science Edition, 2011, 35(5): 476-479. (in Chinese)

[12] Cheng D J, Xu Y J, Liu X S, Zhao L M, Xiong S D, Zhang Z X. Effect of protein kinase C on the tension of passively sensitized human airway smooth muscle. Journal of Chinese Practical Diagnosis and Therapy, 2014, 28(1): 1057-1059.

[13] 李泽. AMPK活性对宰后羊肉能量代谢和肉质的影响及其机理研究[D]. 呼和浩特: 内蒙古农业大学, 2010.

Li Z. Effect of AMPK activity on energy metabolism and meat quality of postmortem lamb and its mechanism research [D]. Hohhot: Inner Mongolia University, 2010. (in Chinese)

[14] Wu Z Z, Liu M. Aaasy of PKC activity and PKA activity in human thyroid cancer. China Journal of Modern Medicine, 2005, 15(2): 195-198.

[15] Culler R D, Smith G C, Cross H R. Relationship of myofibril fragmentation index to certain chemical, physical and sensory characteristics of bovine longissimus muscle. Journal of Food Science, 1978, 43: 1177-1180.

[16] Prates M J A, Costa F J S G E, Ribeiro A M R, Correia A A D. Contribution of major structural changes in myofibrils to rabbit meat tenderisation during ageing. Meat Science, 2002, 61: 103-113.

[17] Lametsch R, Kristensen L, Larsen M R, Therkildsen M, Oksbjerg N, Ertbjerg P. Changes in the muscle proteome after compensatory growth in pigs. Journal of Animal Science, 2006, 84: 918-924.

[18] Huang H G, Larsen M R, Karlsson A H, Pomponio L, Costa LN, Lametsch R. Gel-based phosphoproteomics analysis of sarcoplasmic proteins in postmortem porcine muscle with pH decline rate and time differences. Proteomics, 2011, 11(20): 4063-4076.

[19] Huang H G, Larsen M R, Lametsch R. Changes in phosphorylation of myofibrillar proteins during postmortem development of porcine muscle. Food Chemistry, 2012, 134(3): 1999-2006.

[20] Li C B, Li J, Zhou G H, Ertbjerg P, Brüggemann D A, Huang H G, Karlsson A H, Hviid M, Lundström K. Electrical stimulation affects metabolic enzyme phosphorylation, protease activation, and meat tenderization in beef. Journal of Animal Science, 2012, 90(5): 1638-1649.

[21] Chen L J, Li X, Ni N, Liu Y, Chen L, Wang Z Y, Shen Q W, Zhang D Q.Phosphorylation of myofibrillar proteins in postmortem ovine muscle with different tenderness. Journal of the Science of Food and Agriculture, 2015. doi: 10.1002/jsfa.7244.

[22] 尹靖东. 动物肌肉生物学与肉品科学. 北京: 中国农业大学出版社, 2011: 63-70.

Yin J D. Animal Muscle Biology and Meat Quality. Beijing: China Agricultural University Press, 2011: 63-70. (in Chinese)

[23] Ahmed S H, Lindsey M L. Titin phosphorylation myocardial passive stiffness regulated by the intracellular giant. Circulation Research, 2009, 105: 611-613.

[24] Wang L, Sadayappan S, Kawai M. Cardiac myosin binding protein C phosphorylation affects cross-bridge cycle’s elementary steps in a site-specific manner. Plos One, 2014, 9(11): e113417.

[25] Levine R, Weisberg A, Kulikovskaya I, McClellan G, Winegrad S. Multiple structures of thick filaments in resting cardiac muscle and their influence on cross-bridge interactions. Biophysical Journal, 2001, 81(2): 1070-1082.

[26] Wu S C, Solaro R J. Protein kinase C zeta: A novel regulator of both phosphorylation and de-phosphorylation of cardiac sarcomeric proteins. Journal of Biology Chemistry, 2007, 282(42): 30691-30698.

[27] Monasky M M, Biesiadecki J B, Janssen M L. Increased phosphorylation of tropomyosin, troponin I, and myosin light chain-2 after stretch in rabbit ventricular myocardium under physiological conditions. Journal of Molecular and Cellular Cardiology, 2010, 48: 1023-1028.

[28] England P J. Correlation between contraction and phosphorylation of the inhibitory subunit of troponin in perfused rat heart. Febs Letters, 1975, 50(1): 57-60.

[29] Mark H, Begin K J, Gorga J A, Fishbaugher D E, Lewinter M M, Peter V B. Protein kinase A mediated modulation of acto-myosin kinetics. Journal of Molecular & Cellular Cardiology, 2005, 38(1): 119-125.

[30] 黄明, 薛梅, 黄继超, 徐宝才，郭琳, 徐幸莲, 周光宏. 鸡肉钙激活酶与其它畜禽肉中钙激活酶对钙离子浓度敏感性的比较. 中国农业科学, 2011, 44(12): 2538-2544.

Huang M, Xue M, Huang J C, Xu B C, Guo L, Xu X L, Zhou G H. Calpains in chicken muscle and comparison of calcium sensitivity with other animals. Scientia Agricultura Sinica, 2011, 44(12): 2538-2544. (in Chinese)

[31] Klug G A, Botterman B R, Stull J T. The effect of low-frequency stimulation on myosin light chain phosphorylation in skeletal-muscle. Journal of Biological Chemistry, 1982, 257: 4688-4690.

[32] Di Lisa F, De Tullio R, Salamino F, Barbato R, Melloni E, Siliprandi N, Schiaffino S, Pontremoli S. Specific degradation of troponin T and I by mu-calpain and its modulation by substrate phosphorylation. Biochemical Journal, 1995, 308: 57-61.

[33] Zhang Z, Lawrence J, Stracher A. Phosphorylation of platelet actin binding protein protects against proteolysis by calcium-dependant sulfhydryl protease. Biochemical and Biophysical Research Communications, 1995, 151: 355-360.

[34] D'Alessandro A, Zolla L. Meat science: From proteomics to integrated omics towards system biology. Journal of Proteomics, 2013, 78: 558-577.

[35] 贾文婷. 宰后不同处理对羊肉品质影响的研究[D]. 新疆石河子: 石河子大学, 2013.

Jia W T. Research of the effects of different processing on mutton quality after slaughter [D]. Shihezi, Xinjiang: Shihezi University, 2013. (in Chinese)

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