Chen J W, Evans B R, Yang S H, Teplow D B, Jayaram M
Department of Microbiology, University of Texas, Austin 78712.
Proc Natl Acad Sci U S A. 1991 Jul 15;88(14):5944-8. doi: 10.1073/pnas.88.14.5944.
Binding of a partial proteolytic digest by V8 enzyme of the yeast site-specific recombinase Flp to its target site gives rise to DNA-protein complexes that are smaller than those produced by the full-sized protein. The smallest of these complexes (occupancy of one peptide monomer per site) contains either one of two polypeptides (32 and 28 kDa) of the V8 digestion mixture. The amino termini of both polypeptides map to Ser-129 of Flp, corresponding to V8 cleavage at Glu-128. The relative mobilities of the complexes formed by the V8 peptides indicate that they lack the sharp substrate bend that is characteristic of Flp-derived complexes. A hybrid protein consisting of the amino-terminal one-third of the R recombinase (from Zygosaccharomyces rouxii) and the carboxyl-terminal two-thirds of Flp recognizes the Flp target site.
酵母位点特异性重组酶Flp经V8酶部分酶解后的产物与其靶位点结合，形成的DNA-蛋白质复合物比全长蛋白质形成的复合物小。这些复合物中最小的（每个位点占据一个肽单体）包含V8酶解混合物中的两种多肽（32 kDa和28 kDa）之一。两种多肽的氨基末端均定位到Flp的Ser-129，对应于V8酶在Glu-128处的切割。V8肽形成的复合物的相对迁移率表明，它们缺乏Flp衍生复合物特有的明显底物弯曲。一种由R重组酶（来自鲁氏接合酵母）的氨基末端三分之一和Flp的羧基末端三分之二组成的杂合蛋白可识别Flp靶位点。
