蛋白质磷酸化对肌动球蛋白解离及其乙酰化水平的影响

doi:10.3864/j.issn.0578-1752.2022.07.014

Abstract

Abstract:

【Objective】The objective of this study was to investigate the effects of myosin heavy chain and actin phosphorylation on their acetylation levels, actomyosin dissociation, and ATPase activity, so as to provide a theoretical basis for improving meat tenderness by regulating protein phosphorylation level. 【Method】The homogenate of sheep longissimus dorsi muscle was incubated with alkaline phosphatase inhibitor (inhibiting dephosphorylation) and protein kinase inhibitor (inhibiting phosphorylation) at 4℃ for 0, 0.5, 4, 12, 24, 48, and 72 h to regulate the phosphorylation levels of myosin heavy chain and actin. The protein phosphorylation level was measured by SDS-PAGE and fluorescent staining, and the acetylation level and actomyosin dissociation degree were measured by Western blotting. The ATPase activity was measured using an assay kit. The influence of myosin heavy chain and actin phosphorylation on the structure of actomyosin was analyzed by molecular dynamics simulation. 【Result】The phosphorylation level of myosin heavy chain in the alkaline phosphatase inhibitor treatment group was significantly higher than that in the control and protein kinase inhibitor treatment groups at 4, 12, and 72 h of incubation (P<0.05). The phosphorylation level of actin was significantly higher than that in the control and protein kinase inhibition treatment groups at 4, 12, 24, 48, and 72 h of incubation (P<0.05), which indicated that alkaline phosphatase inhibitors could inhibit the dephosphorylation of myosin heavy chain and actin during incubation in vitro. The acetylation level of actin in the alkaline phosphatase inhibitor treatment group was significantly lower than that in the protein kinase inhibitor treatment group after incubation for 4, 12, 24, 48, and 72 h (P<0.05), while the acetylation level of myosin heavy chain changed irregularly. The results indicated that the phosphorylation of actin inhibited its acetylation, while the phosphorylation of myosin heavy chain had no obvious regularity on its acetylation. The results of molecular dynamics showed that the phosphorylation of the 2nd, 3rd and 54th serine positions of the myosin heavy chain and the 54th and 55th tyrosine positions of actin increased the total energy, potential energy, and kinetic energy of actomyosin. However, the bond energy of actomyosin was reduced, which caused the unstable structure of actomyosin. The dissociation degree of actomyosin in the alkaline phosphatase inhibitor treatment group was always higher than that of the protein kinase inhibitor treatment group during 0-72 h incubation (P<0.05). The ATPase activity was always lower than that in the protein kinase inhibitor treatment group during 0-72 h incubation (P<0.05). The myosin heavy chain and actin phosphorylation promoted actomyosin dissociation. 【Conclusion】The phosphorylation of myosin heavy chain directly promoted the dissociation of actomyosin, while the phosphorylation of actin promoted the dissociation of actomyosin by inhibiting its acetylation.

Key words: phosphorylation, acetylation, myosin heavy chain, actin, dissociation

ZHANG YeJun,ZHANG DeQuan,HOU ChengLi,BAI YuQiang,REN Chi,WANG Xu,LI Xin. Effects of Protein Phosphorylation on the Dissociation and Acetylation Level of Actomyosin[J].Scientia Agricultura Sinica, 2022, 55(7): 1433-1444.

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References 45

[1]	HOLMAN B W B, DAMIAN C, KILGANNON A K, HOPKINS D L. Using shear force, sarcomere length, particle size, collagen content, and protein solubility metrics to predict consumer acceptance of aged beef tenderness. Journal of Texture Studies, 2020, 51(4):559-566. doi: 10.1111/jtxs.12523. doi: 10.1111/jtxs.12523
[2]	CASSENS A M, ARNOLD A N, MILLER R K, GEHRING K B, SAVELL J W. Impact of elevated aging temperatures on retail display, tenderness, and consumer acceptability of beef. Meat Science, 2018, 146:1-8. doi: 10.1016/j.meatsci.2018.07.024. doi: 10.1016/j.meatsci.2018.07.024
[3]	TAYLOR R G, GEESINK G H, THOMPSON V F, KOOHMARAIE M, GOLL D E. Is Z-disk degradation responsible for postmortem tenderization? Journal of Animal Science, 1995, 73(5):1351-1367. doi: 10.2527/1995.7351351x. doi: 10.2527/1995.7351351x
[4]	WANG D Y, ZHANG M H, DENG S Y, XU W M, LIU Y, GENG Z M, SUN C, BIAN H, LIU F. Postmortem changes in actomyosin dissociation, myofibril fragmentation and endogenous enzyme activities of grass carp (Ctenopharyngodon idellus) muscle. Food Chemistry, 2016, 197:340-344. doi: 10.1016/j.foodchem.2015.10.132
[5]	OKITANI A, ICHINOSE N, KOZA M, YAMANAKA K, MIGITA K, MATSUISHI M. AMP and IMP dissociate actomyosin into actin and myosin. Bioscience, Biotechnology, and Biochemistry, 2008, 72(8):2005-2011. doi: 10.1271/bbb.80128. doi: 10.1271/bbb.80128
[6]	OKITANI A, ICHINOSE N, ITOH J, TSUJI Y, ONEDA Y, HATAE K, MIGITA K, MATSUISHI M. Liberation of actin from actomyosin in meats heated to 65℃. Meat Science, 2009, 81(3):446-450. doi: 10.1016/j.meatsci.2008.09.008. doi: 10.1016/j.meatsci.2008.09.008
[7]	BHAT Z F, MORTON J D, MASON S L, BEKHIT A E D A. Role of calpain system in meat tenderness: A review. Food Science and Human Wellness, 2018, 7(3):196-204. doi: 10.1016/j.fshw.2018.08.002. doi: 10.1016/j.fshw.2018.08.002
[8]	PERRIE W T, SMILLIE L B, PERRY S V. A phosphorylated light chain component of myosin from skeletal muscle. Cold Spring Harbor Symposia on Quantitative Biology, 1973, 37:17-18. doi: 10.1101/sqb.1973.037.01.006. doi: 10.1101/sqb.1973.037.01.006
[9]	ALAMO L, WRIGGERS W, PINTO A, BÁRTOLI F, SALAZAR L, ZHAO F Q, CRAIG R, PADRÓN R. Three-dimensional reconstruction of tarantula myosin filaments suggests how phosphorylation may regulate myosin activity. Journal of Molecular Biology, 2008, 384(4):780-797. doi: 10.1016/j.jmb.2008.10.013. doi: 10.1016/j.jmb.2008.10.013
[10]	BRITO R, ALAMO L, LUNDBERG U, GUERRERO J R, PINTO A, SULBARÁN G, GAWINOWICZ M A, CRAIG R, PADRÓN R. A molecular model of phosphorylation-based activation and potentiation of tarantula muscle thick filaments. Journal of Molecular Biology, 2011, 414(1):44-61. doi: 10.1016/j.jmb.2011.09.017. doi: 10.1016/j.jmb.2011.09.017
[11]	KASZA K E, FARRELL D L, ZALLEN J A. Spatiotemporal control of epithelial remodeling by regulated myosin phosphorylation. PNAS, 2014, 111(32):11732-11737. doi: 10.1073/pnas.1400520111. doi: 10.1073/pnas.1400520111
[12]	陈立娟, 李欣, 李铮, 李培迪, 李仲文, 张德权. 蛋白质磷酸化调控羊肉肌原纤维蛋白的功能. 中国农业科学, 2016, 49(7):1360-1370.
	CHEN L J, LI X, LI Z, LI P D, LI Z W, ZHANG D Q. Protein phosphorylation on the function of myofibrillar proteins in mutton muscle. Scientia Agricultura Sinica, 2016, 49(7):1360-1370. (in Chinese)
[13]	张艳, 李欣, 李铮, 李蒙, 刘永峰, 张德权. 冰温贮藏对羊肉中蛋白质磷酸化水平的影响. 中国农业科学, 2016, 49(22):4429-4440. doi: 10.3864/j.issn.0578-1752.2016.22.015. doi: 10.3864/j.issn.0578-1752.2016.22.015
	ZHANG Y, LI X, LI Z, LI M, LIU Y F, ZHANG D Q. Effects of controlled freezing point storage on the protein phosphorylation level. Scientia Agricultura Sinica, 2016, 49(22):4429-4440. doi: 10.3864/j.issn.0578-1752.2016.22.015. (in Chinese) doi: 10.3864/j.issn.0578-1752.2016.22.015
[14]	李蒙, 李铮, 李欣, 杜曼婷, 宋璇, 张德权. 磷酸化水平对肌红蛋白稳定性的影响. 中国农业科学, 2017, 50(22):4382-4388. doi: 10.3864/j.issn.0578-1752.2017.22.014. doi: 10.3864/j.issn.0578-1752.2017.22.014
	LI M, LI Z, LI X, DU M T, SONG X, ZHANG D Q. Effect of phosphorylation level on myoglobin stability. Scientia Agricultura Sinica, 2017, 50(22):4382-4388. doi: 10.3864/j.issn.0578-1752.2017.22.014. (in Chinese) doi: 10.3864/j.issn.0578-1752.2017.22.014
[15]	高星, 李欣, 李铮, 丁武, 张德权. 肌动球蛋白磷酸化对其解离的影响. 食品科学, 2017, 38(9):21-26. doi: 10.7506/spkx1002-6630-201709004. doi: 10.7506/spkx1002-6630-201709004
	GAO X, LI X, LI Z, DING W, ZHANG D Q. Effect of phosphorylation on actomyosin dissociation. Food Science, 2017, 38(9):21-26. doi: 10.7506/spkx1002-6630-201709004. (in Chinese) doi: 10.7506/spkx1002-6630-201709004
[16]	GAO X, LI X, LI Z, DU M T, ZHANG D Q. Dephosphorylation of myosin regulatory light chain modulates actin-myosin interaction adverse to meat tenderness. International Journal of Food Science and Technology, 2017, 52(6):1400-1407. doi: 10.1111/ijfs.13343
[17]	JIANG S W, LIU Y S, SHEN Z L, ZHOU B, SHEN Q W. Acetylome profiling reveals extensive involvement of lysine acetylation in the conversion of muscle to meat. Journal of Proteomics, 2019, 205:103412. doi: 10.1016/j.jprot.2019.103412. doi: 10.1016/j.jprot.2019.103412
[18]	ZHOU B, SHEN Z L, LIU Y S, WANG C T, SHEN Q W. Proteomic analysis reveals that lysine acetylation mediates the effect of antemortem stress on postmortem meat quality development. Food Chemistry, 2019, 293:396-407. doi: 10.1016/j.foodchem.2019.04.122. doi: 10.1016/j.foodchem.2019.04.122
[19]	HOFMANN T G, MÖLLER A, SIRMA H, ZENTGRAF H, TAYA Y, DRÖGE W, WILL H, SCHMITZ M L. Regulation of p53 activity by its interaction with homeodomain-interacting protein kinase-2. Nature Cell Biology, 2002, 4(1):1-10. doi: 10.1038/ncb715. doi: 10.1038/ncb715
[20]	LI Z, BRIDGES B, OLSON J, WEINMAN S A. The interaction between acetylation and serine-574 phosphorylation regulates the apoptotic function of FOXO3. Oncogene, 2017, 36(13):1887-1898. doi: 10.1038/onc.2016.359. doi: 10.1038/onc.2016.359
[21]	陈立娟, 李欣, 杨扬, 陈丽, 倪娜, 张德权. 不同嫩度羊肉肌浆蛋白质磷酸化水平随宰后成熟时间变化的研究. 现代食品科技, 2015, 31(4):95-101. doi: 10.13982/j.mfst.1673-9078.2015.4.016. doi: 10.13982/j.mfst.1673-9078.2015.4.016
	CHEN L J, LI X, YANG Y, CHEN L, NI N, ZHANG D Q. Analyzing the changes in sarcoplasmic protein phosphorylation with respect to postmortem ageing times in mutton with different levels of tenderness. Modern Food Science and Technology, 2015, 31(4):95-101. doi: 10.13982/j.mfst.1673-9078.2015.4.016. (in Chinese) doi: 10.13982/j.mfst.1673-9078.2015.4.016
[22]	高星, 李欣, 李铮, 杜曼婷, 张彩霞, 张德权, 丁武. 宰后肌肉中肌球蛋白磷酸化调控肌动球蛋白解离作用机制. 中国农业科学, 2016, 49(16):3199-3207. doi: 10.3864/j.issn.0578-1752.2016.16.013. doi: 10.3864/j.issn.0578-1752.2016.16.013
	GAO X, LI X, LI Z, DU M T, ZHANG C X, ZHANG D Q, DING W. The mechanism of myosin phosphorylation regulating actomyosin dissociation of skeletal muscle during postmortem. Scientia Agricultura Sinica, 2016, 49(16):3199-3207. doi: 10.3864/j.issn.0578-1752.2016.16.013. (in Chinese) doi: 10.3864/j.issn.0578-1752.2016.16.013
[23]	CHEN X R, WANG X T, HAO M Q, ZHOU Y H, CUI W Q, XING X X, XU C G, BAI J W, LI Y H. Homology modeling and virtual screening to discover potent inhibitors targeting the imidazole glycerophosphate dehydratase protein in Staphylococcus xylosus. Frontiers in Chemistry, 2017, 5:98. doi: 10.3389/fchem.2017.00098. doi: 10.3389/fchem.2017.00098
[24]	LIU M S, WEI Y C, LI X, QUEK S Y, ZHAO J, ZHONG H Z, ZHANG D Q, LIU Y F. Quantitative phosphoproteomic analysis of caprine muscle with high and low meat quality. Meat Science, 2018, 141:103-111. doi: 10.1016/j.meatsci.2018.01.001. doi: 10.1016/j.meatsci.2018.01.001
[25]	SCHIAFFINO S, REGGIANI C. Fiber types in mammalian skeletal muscles. Physiological Reviews, 2011, 91(4):1447-1531. doi: 10.1152/ physrev.00031.2010. doi: 10.1152/ physrev.00031.2010
[26]	KARLSSON A H, KLONT R E, FERNANDEZ X. Skeletal muscle fibres as factors for pork quality. Livestock Production Science, 1999, 60(2/3):255-269. doi: 10.1016/S0301-6226(99)00098-6. doi: 10.1016/S0301-6226(99)00098-6
[27]	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.
	YIN J D. Animal Muscle Biology and Meat Quality. Beijing: China Agricultural University Press, 2011. (in Chinese)
[28]	李胜杰, 徐幸莲, 周光宏. 宰后肌动球蛋白解离对肉品嫩度的影响研究进展. 食品科学, 2010, 31(21):442-445.
	LI S J, XU X L, ZHOU G H. Research advances in the influence of actomyosin dissociation on postharvest meat tenderness. Food Science, 2010, 31(21):442-445. (in Chinese)
[29]	EGELHOFF T T, LEE R J, SPUDICH J A. Dictyostelium myosin heavy chain phosphorylation sites regulate myosin filament assembly and localization in vivo. Cell, 1993, 75(2):363-371. doi: 10.1016/0092-8674(93)80077-R
[30]	NORWOOD TORO L E, WANG Y R, CONDEELIS J S, JONES J G, BACKER J M, BRESNICK A R. Myosin-IIA heavy chain phosphorylation on S1943 regulates tumor metastasis. Experimental Cell Research, 2018, 370(2):273-282. doi: 10.1016/j.yexcr.2018.06.028. doi: 10.1016/j.yexcr.2018.06.028
[31]	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 post-mortem ovine muscle with different tenderness. Journal of the Science of Food and Agriculture, 2016, 96(5):1474-1483. doi: 10.1002/jsfa.7244. doi: 10.1002/jsfa.7244
[32]	LIU X, SHU S, HONG M S S, LEVINE R L, KORN E D. Phosphorylation of actin Tyr-53 inhibits filament nucleation and elongation and destabilizes filaments. Proceedings of the National Academy of Sciences of the United States of America, 2006, 103(37):13694-13699.
[33]	GANNON J, STAUNTON L, O'CONNELL K, DORAN P, OHLENDIECK K. Phosphoproteomic analysis of aged skeletal muscle. International Journal of Molecular Medicine, 2008, 22(1):33-42.
[34]	PAPAKONSTANTI E A, STOURNARAS C. Association of PI-3 kinase with PAK1 leads to actin phosphorylation and cytoskeletal reorganization. Molecular Biology of the Cell, 2002, 13(8):2946-2962. doi: 10.1091/mbc.02-01-0599. doi: 10.1091/mbc.02-01-0599
[35]	CARLIER M F, PANTALONI D. Control of actin dynamics in cell motility. Journal of Molecular Biology, 1997, 269(4):459-467. doi: 10.1006/jmbi.1997.1062. doi: 10.1006/jmbi.1997.1062
[36]	HOWARD P K, SEFTON B M, FIRTEL R A. Tyrosine phosphorylation of actin in dictystelium associated with cell-shape changes. Science, 1993, 259(5092):241-244. doi: 10.1126/science.7678470
[37]	KARPLUS M. Molecular dynamics simulations of biomolecules. Accounts of Chemical Research, 2002, 35(6):321-323. doi: 10.1021/ar020082r. doi: 10.1021/ar020082r
[38]	ZHANG Y J, LI X, ZHANG D Q, REN C, BAI Y Q, IJAZ M, WANG X, ZHAO Y X. Acetylation of sarcoplasmic and myofibrillar proteins were associated with ovine meat quality attributes at early postmortem. Food Science of Animal Resources, 2021, 41(4):650-663. doi: 10.5851/kosfa.2021.e22. doi: 10.5851/kosfa.2021.e22
[39]	HABIBIAN J, FERGUSON B S. The crosstalk between acetylation and phosphorylation: emerging new roles for HDAC inhibitors in the heart. International Journal of Molecular Sciences, 2018, 20(1):102. doi: 10.3390/ijms20010102. doi: 10.3390/ijms20010102
[40]	SAMANT S A, PILLAI V B, SUNDARESAN N R, SHROFF S G, GUPTA M P. Histone deacetylase 3 (HDAC3)-dependent reversible lysine acetylation of cardiac myosin heavy chain isoforms modulates their enzymatic and motor activity. The Journal of Biological Chemistry, 2015, 290(25):15559-15569. doi: 10.1074/jbc.M115.653048. doi: 10.1074/jbc.M115.653048
[41]	VISWANATHAN M C, BLICE-BAUM A C, SCHMIDT W, FOSTER D B, CAMMARATO A. Pseudo-acetylation of K326 and K328 of actin disrupts Drosophila melanogaster indirect flight muscle structure and performance. Frontiers in Physiology, 2015, 6:116. doi: 10.3389/fphys.2015.00116. doi: 10.3389/fphys.2015.00116
[42]	SCHMIDT W, VISWANATHAN M, FOSTER D B, CAMMARATO A. Acetylation of k326 and k328 on actin boosts contractile properties of muscle in vitro and in vivo. Biophysical Journal, 2017, 112(3):483a.
[43]	SCHMIDT W, VISWANATHAN M, BLICE-BAUM A C, FOSTER D B, CAMMARATO A. Pseudo-acetylation of actin residues k326 and k328 disrupts drosophila flight performance and muscle structure. Biophysical Journal, 2015, 108(2):421a-422a.
[44]	SCHMIDT W M, FOSTER D B, CAMMARATO A. Acetylation of actin k328 contributes to a loss in tropomyosin-mediated inhibition of myosin binding. Biophysical Journal, 2019, 116(3):457a.
[45]	ZHANG Y J, LI X, ZHANG D Q, BAI Y Q, WANG X. Effects of acetylation on dissociation and phosphorylation of actomyosin in postmortem ovine muscle during incubation at 4℃ in vitro. Food Chemistry, 2021, 356:129696. doi: 10.1016/j.foodchem.2021.129696. doi: 10.1016/j.foodchem.2021.129696

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蛋白 Protein	磷酸化位点 Location of phosphorylation sites
肌动蛋白 Actin	S54, Y55, S62, Y93, T150, T151, Y168, Y171, Y200, T231, S234, S235, S236, S237, S241, T251, S325, S370
肌球蛋白重链 Myosin heavy chain	S2, S3, S54, T257, T258, T381, Y389, T415, Y424, T452, Y556, T684, S732, S742, T790, S814, Y820, S846, T885, S897, T915, S952, T964, T983, T992, T997, T1023, T1025, T1029, S1041

	总能量 Total energy (kcal/mol)	势能 Potential energy (kcal/mol)	动能 Kinetic energy (kcal/mol)	键能 Bond energy (kcal/mol)
未磷酸化肌动球蛋白 Unphosphorylated actomyosin	-362733.033	-453252.402	90519.369	4563.800
磷酸化肌动球蛋白 Phosphorylated actomyosin	-405716.468	-504188.345	98471.877	4509.000

Effects of Protein Phosphorylation on the Dissociation and Acetylation Level of Actomyosin

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