ISOLATION OF PEPTIDES THAT INHIBIT BINDING OF BASIC FIBROBLAST GROWTH-FACTOR TO ITS RECEPTOR FROM A RANDOM PHAGE-EPITOPE LIBRARY
ISOLATION OF PEPTIDES THAT INHIBIT BINDING OF BASIC FIBROBLAST GROWTH-FACTOR TO ITS RECEPTOR FROM A RANDOM PHAGE-EPITOPE LIBRARY
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DOI：
10.1073/pnas.90.22.10643
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发表时间：
1993-11-15
影响因子：
11.1
通讯作者：
KATCHALSKIKATZIR, E
中科院分区：
文献类型：
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作者：
YAYON, A;AVIEZER, D;KATCHALSKIKATZIR, E
已知碱性成纤维细胞生长因子（bFGF）以高亲和力和肝素依赖性方式与其细胞表面受体结合。为了预测bFGF上的受体识别位点，我们用单克隆抗体DG 2和DE 6筛选了噬菌体-表位文库，它们抑制bFGF与其受体的结合。在亲和分离的cDNA上，我们确定了几个肽序列作为bFGF上推定的抗体结合表位。所鉴定的文库表位共有序列Pro-（Pro/Ser）-Gly-His-（Tyr/Phe）-Lys，分别对应于bFGF的两个连续蛋白序列：Pro-Pro-Gly-His-Phe-Lys和Arg-Thr-Gly-Gln-Tyr-Lys，位于bFGF的13-18和120-125位氨基酸。相应的噬菌体表位或上述bFGF序列的合成肽特异性地抑制抗体与bFGF的结合，阻断bFGF与其高亲和力受体的结合，并在亚微摩尔肽浓度下抑制血管内皮细胞的基础和bFGF诱导的增殖。由抗体识别的肽对bFGF结合和生物活性的有效抑制表明，这些序列在功能上参与受体结合，并且可能构成bFGF上的受体结合决定簇的一部分。
Basic fibroblast growth factor (bFGF) is known to bind to its cell-surface receptors with high affinity and in a heparin-dependent manner. In an attempt to predict the receptor recognition site on bFGF we screened phage-epitope libraries with monoclonal antibodies DG2 and DE6, which inhibit bFGF binding to its receptor. On the affinity-isolated phages, we identified several peptide sequences as the putative antibody-binding epitopes on bFGF. The identified library epitopes shared the consensus sequence Pro-(Pro/Ser)-Gly-His-(Tyr/Phe)-Lys, corresponding to two continuous protein sequences of bFGF: Pro-Pro-Gly-His-Phe-Lys and Arg-Thr-Gly-Gln-Tyr-Lys at amino acids 13-18 and 120-125 of bFGF, respectively. Synthetic peptides of the corresponding phage epitopes or of the above bFGF sequences specifically inhibited binding of the antibodies to bFGF, blocked binding of bFGF to its high-affinity receptor, and inhibited basal and bFGF-induced proliferation of vascular endothelial cells at submicromolar peptide concentrations. The potent inhibition of bFGF binding and biological activity by peptides recognized by the antibodies suggests that these sequences are functionally involved in receptor binding and may constitute part of the receptor-binding determinants on bFGF.