Zagotta M T, Wilson D B
Section of Biochemistry, Molecular and Cell Biology, Cornell University, Ithaca, New York 14853.
J Bacteriol. 1990 Feb;172(2):912-21. doi: 10.1128/jb.172.2.912-921.1990.
Western blot (immunoblot) analysis of cell extracts from induced bacteriophage lambda lysogens probed with S-protein-specific antibody (raised against an S--beta-galactosidase fusion protein) demonstrated that the bacteriophage lambda S protein begins to appear 10 min after phage induction and is localized to the inner membrane at all times during the lytic cycle. Between 100 and 1,000 molecules of S protein per cell were present at the time of phage-induced lysis. Western blots of chemically cross-linked membranes from induced lysogens showed a ladder of bands at 18, 24, 32, and 42 kilodaltons (the S-protein monomer ran at 8 kilodaltons) that reacted with anti-S-protein antibody. Thus, the S protein appears to reside in the inner membrane as a multimer, and the molecular weights of the cross-linked species are consistent with those of S-protein homopolymers. Sodium dodecyl sulfate-resistant dimers were also detected when S protein was purified by immunoprecipitation.
用S蛋白特异性抗体(针对S-β-半乳糖苷酶融合蛋白产生)对诱导的噬菌体λ溶原菌的细胞提取物进行蛋白质免疫印迹(免疫印迹)分析表明,噬菌体λ S蛋白在噬菌体诱导后10分钟开始出现,并在裂解周期的所有时间都定位于内膜。在噬菌体诱导裂解时,每个细胞存在100至1000个S蛋白分子。对诱导溶原菌的化学交联膜进行的蛋白质免疫印迹显示,在18、24、32和42千道尔顿处有一系列条带(S蛋白单体的分子量为8千道尔顿)与抗S蛋白抗体发生反应。因此,S蛋白似乎以多聚体形式存在于内膜中,交联物种的分子量与S蛋白同聚物的分子量一致。通过免疫沉淀纯化S蛋白时,还检测到了耐十二烷基硫酸钠的二聚体。