兔前顶体蛋白原及一种新型前顶体蛋白原相关cDNA的cDNA克隆与测序

Cloning and sequencing of cDNAs for rabbit preproacrosin and a novel preproacrosin-related cDNA.

作者信息

Richardson R T, O'Rand M G

机构信息

Department of Cell Biology and Anatomy, University of North Carolina at Chapel Hill 27599.

出版信息

Biochim Biophys Acta. 1994 Sep 13;1219(1):215-8. doi: 10.1016/0167-4781(94)90275-5.

DOI:10.1016/0167-4781(94)90275-5

PMID:8086468

Abstract

A 1414 bp cDNA for rabbit preproacrosin (RPA) and a related short preproacrosin (shRPA) cDNA of 951 bp were cloned and sequenced. RPA's 431 amino acid open reading frame encodes a 46,422 kDa protein. shRPA is identical to RPA except that it lacks an internal stretch of 468 bp, such that the encoded protein has a deduced molecular mass of 29,965 kDa. Antiserum against a synthetic peptide representing the light chain of rabbit proacrosin was used on Western blots of rabbit testis and sperm. Under reducing conditions, it revealed two major groups of bands at 50-57 and 29-32 kDa. Several lines of evidence suggest that shRPA is a splice variant of proacrosin and that it encodes a 30-33 kDa protein similar to sperminogen (Siegel, M. et al. (1987) Biol. Reprod. 36, 1063-1068), but apparently lacking proteinase activity.

摘要

克隆并测序了兔前体顶体蛋白酶（RPA）的1414 bp cDNA以及相关的951 bp短前体顶体蛋白酶（shRPA）cDNA。RPA的431个氨基酸开放阅读框编码一种46.422 kDa的蛋白质。shRPA与RPA相同，只是它缺少一段468 bp的内部片段，因此编码的蛋白质推导分子量为29.965 kDa。用针对代表兔前体顶体蛋白酶轻链的合成肽的抗血清对兔睾丸和精子进行蛋白质免疫印迹分析。在还原条件下，它在50 - 57 kDa和29 - 32 kDa处显示出两组主要条带。几条证据表明，shRPA是前体顶体蛋白酶的剪接变体，它编码一种类似于精子原蛋白的30 - 33 kDa蛋白质（Siegel, M.等人（1987年）《生物繁殖》36卷，1063 - 1068页），但显然缺乏蛋白酶活性。