Orbitrap Fusion Lumos三合一质谱技术分析鳜鱼、金鲳鱼和鲟鱼肌动蛋白肽指纹图谱的差异性

Differences in Actin Peptide Fingerprints of Mandarin Fish,Golden Pomfret and Sturgeon Analyzed by Orbitrap Fusion Lumos Mass Spectrometry

利用Orbitrap Fusion Lumos三合一质谱技术分析鳜鱼、金鲳鱼和鲟鱼肌动蛋白的肽指纹图谱,阐明3种鱼类肌动蛋白的氨基酸序列差异性及特征肽段。结果表明,鳜鱼肌动蛋白特征酶解肽段鉴定为骨骼肌α-肌动蛋白、金鲳鱼和鲟鱼都为β-肌动蛋白。鳜鱼肌动蛋白酶解后可检测到29个肽段;金鲳鱼肌动蛋白检测到24个肽段,且全为特征肽段;鲟鱼肌动蛋白检出13个肽段。与另外2种鱼类肌动蛋白的肽质指纹图谱相比,鳜鱼骨骼肌α-肌动蛋白有10个差异肽段,金鲳鱼β-肌动蛋白有5个差异肽段,鲟鱼β-肌动蛋白仅有3个差异肽段。从氨基酸全序列的排列方式来看,鳜鱼骨骼肌α-肌动蛋白与金鲳鱼β-肌动蛋白从相似序列开始有21处氨基酸变异,与鲟鱼β-肌动蛋白从相似序列开始有17处氨基酸变异,鲟鱼β-肌动蛋白的氨基酸序列与金鲳鱼β-肌动蛋白的序列同源性较高,仅有3处氨基酸变异,本研究将为名贵鱼类质量品质属性的鉴定及鱼类肌动蛋白标准品的研制提供基础,为鱼类蛋白功能产品开发利用及快速定量检测提供理论依据。

The peptide mass fingerprints of actin from Siniperca chuatsi,Trachinotus ovatus and Acipenser sinensis were analyzed by Orbitrap Fusion Lumos mass spectrometry.The differences in the amino acid sequence and the signature peptides of actin from the three fishes were clarified.Results showed that the signature peptides were identified in the enzymatic hydrolysate of skeletal muscleα-actin for S.chuatsi,while the signature peptides were derived fromβ-actin for both T.ovatus and A.sinensis.Totally 29 peptides were detected in S.chuatsi actin,24 peptides in T.ovatus actin,all being the signature peptides,and 13 peptides in A.sinensis actin.Compared with the actin fingerprints of the two other fishes,10 differential peptides were identified in skeletal muscleα-actin of S.chuatsi,five differential peptides inβ-actin of T.ovatus,and only three differential peptides inβ-actin of A.sinensis.According to the arrangement of amino acid sequences,there were 21 amino acid substitutions between S.chuatsi skeletal muscleα-actin and T.ovatusβ-actin,starting from the similar sequences,and 17 amino acid substitutions between S.chuatsi skeletal muscleα-actin and A.sinensisβ-actin.The amino acid sequences of A.sinensis and T.ovatusβ-actin showed high homology to each other with only three amino acid substitutions.The results of this study will lay a foundation for the identification of quality properties of rare fish and the development of fish actin standards,and provide a theoretical basis for the development,utilization and rapid quantitative detection of functional fish protein products.