Gårdsvoll H, van Zonneveld A J, Holm A, Eldering E, van Meijer M, Danø K, Pannekoek H
Department of Biochemistry, Academic Medical Center, University of Amsterdam, The Netherlands.
FEBS Lett. 1998 Jul 17;431(2):170-4. doi: 10.1016/s0014-5793(98)00742-x.
Large random hexa- and decapenta-peptide libraries were constructed and displayed on the surface of the filamentous phagemid pComb8. Panning of the hexa-peptide library on immobilized plasminogen activator inhibitor 1 (PAI-1) specifically selected a minor fraction of concatemers, indicating that binding to PAI-1 requires an extended amino acid sequence. Accordingly, the decapenta-peptide library exclusively yielded PAI-1 binding peptides of 15 amino acid residues. None of these phage-bound peptides prevented the interaction between PAI-1 and its target serine protease urokinase (u-PA). To isolate peptides that block the interaction between PAI-1 and u-PA, phages bound to immobilized PAI-1 were eluted by incubation with u-PA. Remarkably, this procedure resulted in elution of a unique phage type that harbors a concatemer of decapentamers, consisting of 49 amino acid residues with no obvious similarity to the primary sequence of PAI-1 or u-PA.
构建了大型随机六肽和十五肽文库，并将其展示在丝状噬菌粒pComb8的表面。在固定化纤溶酶原激活物抑制剂1（PAI-1）上筛选六肽文库，特异性地选择了一小部分串联体，这表明与PAI-1结合需要一个延伸的氨基酸序列。因此，十五肽文库专门产生了15个氨基酸残基的PAI-1结合肽。这些噬菌体结合的肽均未阻止PAI-1与其靶丝氨酸蛋白酶尿激酶（u-PA）之间的相互作用。为了分离阻断PAI-1与u-PA相互作用的肽，将与固定化PAI-1结合的噬菌体与u-PA一起孵育进行洗脱。值得注意的是，这一过程导致洗脱了一种独特的噬菌体类型，其含有十五联体的串联体，由49个氨基酸残基组成，与PAI-1或u-PA的一级序列没有明显相似性。
