超营养补充维生素E影响死后牛肉腰最长肌肌红蛋白翻译后修饰

Yifei Wang, Shuting Li, Jing Chen, Hai-ying Zhu, B. Harsh, D. Boler, A. Dilger, D. Shike, S. Suman
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引用次数: 0

摘要

肌红蛋白(Mb)的翻译后修饰(PTM)可以影响鲜肉的颜色稳定性。膳食补充维生素E通过延缓脂质氧化诱导的Mb氧化和影响死后牛肉骨骼肌蛋白质组谱来改善牛肉颜色稳定性。然而,维生素E对死后牛肉骨骼肌中Mb PTM的影响还有待研究。因此,本研究的目的是研究膳食维生素E对死后牛肉腰最长肌中Mb PTM的影响。采用宰后24 h的9条维生素e (VITE;1000 IU维生素E日粮/小母牛·d - 1, 89 d)和9个对照组(CONT;没有补充维生素E)小母牛。采用十二烷基硫酸钠-聚丙烯酰胺凝胶电泳法对牛腰最长肌其他肌浆蛋白进行分离。串联质谱法鉴定了代表Mb的蛋白带(17 kDa)中的多个PTM(磷酸化、乙酰化、4-羟基壬烯醛烷基化、甲基化、二甲基化、三甲基化和羧甲基化)。易被磷酸化的氨基酸是苏氨酸(T)和酪氨酸(Y),而赖氨酸(K)残基容易被其他PTM所磷酸化。在CONT和VITE样品的Mb中发现了相同的磷酸化位点(T34、T67、Y103)、羧甲基化位点(K77、K78)和4-羟基壬烯醛烷基化位点(K77、K78、K79),表明这些PTM不受补充维生素E的影响。尽管如此,在CONT和VITE样本的Mb中发现了乙酰化、甲基化、二甲基化和三甲基化的差异。总的来说,CONT中修饰的氨基酸数量多于VITE,这表明补充维生素E减少了Mb中翻译后修饰残基的数量。此外,K87、K96、K98和K102的PTM是CONT所特有的,而K118的PTM是VITE所特有的。上述结果提示,在肉牛饲粮中添加维生素E可以保护Mb中氨基酸残基(尤其是靠近近端组氨酸的氨基酸残基)不发生PTM,从而提高Mb的氧化还原稳定性。
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Supranutritional Supplementation of Vitamin E Influences Myoglobin Post-Translational Modifications in Postmortem Beef Longissimus Lumborum Muscle
Post-translational modifications (PTM) in myoglobin (Mb) can influence fresh meat color stability. Dietary supplementation of vitamin E improves beef color stability by delaying lipid oxidation–induced Mb oxidation and influences proteome profile of postmortem beef skeletal muscles. Nonetheless, the influence of vitamin E on Mb PTM in postmortem beef skeletal muscles has yet to be investigated. Therefore, the objective of the current study was to examine the effect of dietary vitamin E on Mb PTM in postmortem beef longissimus lumborum muscle. Beef longissimus lumborum muscle samples (24 h postmortem) were obtained from the carcasses of 9 vitamin E–supplemented (VITE; 1,000 IU vitamin E diet/heifer·d−1for 89 d) and 9 control (CONT; no supplemental vitamin E) heifers. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis was used to separate Mb from other sarcoplasmic proteins of beef longissimus lumborum muscle. Tandem mass spectrometry identified multiple PTM (phosphorylation, acetylation, 4-hydroxynonenalalkylation, methylation, dimethylation, trimethylation, and carboxymethylation) in the protein bands (17 kDa) representing Mb. The amino acids susceptible to phosphorylation were threonine (T) and tyrosine (Y), whereas lysine (K) residues were prone to other PTM. The same sites of phosphorylation (T34, T67, Y103), carboxymethylation (K77, K78), and 4-hydroxynonenal alkylation (K77, K78, K79) were identified in Mb from CONT and VITE samples, indicating that these PTM were not influenced by the vitamin E supplementation in cattle. Nonetheless, differential occurrence of acetylation, methylation, dimethylation, and trimethylation were identified in Mb from CONT and VITE samples. Overall, a greater number of amino acids were modified in CONT than VITE, suggesting that the supplementation of vitamin E decreased thenumbers of post-translationally modified residues in Mb. Additionally, PTM at K87, K96, K98, and K102 were unique to CONT, whereas PTM at K118 were unique to VITE. These findings suggested that dietary supplementation of vitamin E in beef cattle might protect amino acid residues in Mb—especially those located spatially close to proximal histidine—from undergoing PTM, thereby improving Mb redox stability.
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