Kwon B S, Wakulchik M, Liu C C, Persechini P M, Trapani J A, Haq A K, Kim Y, Young J D
Department of Microbiology and Immunology, Indiana Univ. School of Medicine, Indianapolis 46223.
Biochem Biophys Res Commun. 1989 Jan 16;158(1):1-10. doi: 10.1016/s0006-291x(89)80168-8.
Purified murine lymphocyte pore-forming protein (PFP or perforin) was partially sequenced. Oligonucleotides synthesized on the basis of this sequence information were used to screen a murine cytotoxic T lymphocyte (CTL) cDNA library. Seven clones were obtained, two of which were sequenced, providing full-length sequence information on PFP. Murine PFP (534 a.a.) is 68% identical to human PFP. Hydropathic analysis revealed a predominantly hydrophilic protein with some hydrophobic domains, including a region (a.a. 191-251) that could contain putative membrane-spanning domains. PFP is approx. 20% identical to human C7, C8 and C9 within a region encompassing 270 a.a., confirming previous immunological cross-reactivity studies. Northern blot analysis showed that expression of PFP but not of a serine esterase transcript is enhanced in a CTL line by antigen receptor-stimulation. Southern blot analysis of mouse genomic DNA indicated that PFP is encoded as a single-copy gene with the coding region contained within 10 kilobases of genomic DNA.
对纯化的小鼠淋巴细胞穿孔蛋白（PFP或穿孔素）进行了部分测序。根据该序列信息合成的寡核苷酸用于筛选小鼠细胞毒性T淋巴细胞（CTL）cDNA文库。获得了7个克隆，其中2个进行了测序，提供了PFP的全长序列信息。小鼠PFP（534个氨基酸）与人类PFP的同源性为68%。亲水性分析显示，该蛋白主要为亲水性，但含有一些疏水区，包括一个可能包含假定跨膜结构域的区域（氨基酸191 - 251）。在包含270个氨基酸的区域内，PFP与人类C7、C8和C9的同源性约为20%，这证实了先前的免疫交叉反应性研究。Northern印迹分析表明，在CTL系中，抗原受体刺激可增强PFP而非丝氨酸酯酶转录本的表达。对小鼠基因组DNA的Southern印迹分析表明，PFP作为单拷贝基因编码，其编码区包含在10千碱基的基因组DNA内。
