【摘要】 棕榈酰蛋白硫酯酶1(palmitoyl protein thioesterase 1,PPT1)能够催化棕榈酰蛋白的去棕榈酰化修饰,通过调节蛋白的棕榈酰化修饰参与多种细胞生物学进程.从日本七鳃鳗(Lampetra japonica)白细胞cDNA文库的表达序列标签(Expressed Sequence Tag,EST)中获取PPT1基因片段,经RT-PCR扩增获得七鳃鳗PPT1蛋白的全序列cDNA.基因全长为972bp,编码323个氨基酸组成的蛋白质,理论分子量为36.6kDa,等电点为5.91,并含有1个信号肽.通过同源序列比对分析结果显示,日本七鳃鳗PPT1与其他物种PPT具有很高的同源性.利用实时荧光定量PCR法检测到PPT1基因在七鳃鳗的各组织中广泛表达,其中在白细胞、心脏、肠、鳃中表达量较高,在肝脏中表达量最低.本研究为探讨七鳃鳗PPT1功能研究奠定了理论基础.更多还原

【Abstract】 Palmitoyl-protein thioesterase 1(PPT1)is able to hydrolyze thioester linkages in palmitoylproteins,and removes palmitate residues.PPT1 plays an important role in cell growth and development by protein palmitoylation/depalmitoylation reactions.In this study,we found that there is a homology EST sequence tags with PPT1 of the higher vertebrates in leukocytes cDNA library from Lampetra japonica.The whole cDNA of PPT1 was obtained by RT-PCR.The ORF of the Lampetra japonica PPT1(Lj-PPT1)was 972 bp in length,and it encodes a protein of 323 amino acids with a predicted molecular mass of 36.6kDa and a theoretical isoelectric point of 5.91,and the protein contains a signal peptide.Multiple sequence alignments revealed that Lj-PPT1 shares high sequence homology with the PPT1 of other species.Real-time quantitative PCR demonstrated the wide distribution of Lj-PPT1 in lamprey tissues,and the Lj-PPT1 mRNA was present at higher levels in leukocytes,heart,intestines,and gills,and at the lowest level in the liver.The results obtained in the present study will help us to understand the roles of PPT1 in the lamprey.更多还原