Pattern and spacing of basic amino acids in heparin binding sites
Pattern and spacing of basic amino acids in heparin binding sites
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DOI：
10.1006/abbi.1997.0147
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发表时间：
1997-07-01
影响因子：
3.9
通讯作者：
Linhardt, RJ
中科院分区：
文献类型：
--
作者：
Fromm, JR;Hileman, RE;Linhardt, RJ
糖胺聚糖（GAG）-蛋白质相互作用调节无数的生理和病理过程，但缺乏对这些分子如何相互作用的了解，使用肽类似物以及通过检查已知的肝素结合位点研究了肝素结合位点中碱性氨基酸（精氨酸（R）和赖氨酸（K））的模式和间距的作用，具有一般结构RnW（n = 3-9，其中酪氨酸（W））的肽合成了它们与肝素的相互作用，通过等温滴定量热法测定，结合亲和力随着 R 残基数量的增加而增加，R 的 9 聚体 (R9W) 在生理条件下与肝素的结合与酸性成纤维细胞生长因子一样紧密，尽管它们对肝素的亲和力很高，但较长的碱性氨基酸在肝素结合蛋白中并不常见，已知的肝素结合位点最常见包含由一个分隔的单个分离的碱性氨基酸非碱性氨基酸，合成具有结构 H3CCONH-GRRG(m)RRG(5-m)-CONH2（表示为 RRG(m)RR 肽系列）和 H3CCONH-GRRRG(m)RG(5-m)-CONH2（表示为 RRRG(m)R 肽系列）（其中 m = 0-5）的肽，以检验肝素结合位点中碱性氨基酸的间距最佳的假设安排与不同的 GAG 相互作用，通过亲和色谱法测量，当 m = 0 时，-RRG(m)RR- 和 -RRRG(m)R- 肽系列中的肽与肝素结合最紧密。相反，当m = 0或1时，-RRG(m)RR-肽系列与硫酸乙酰肝素相互作用最紧密，而当m = 3时，-RRRG(m)R-肽系列结合最紧密。这些结果与我们对肝素和硫酸乙酰肝素结构的理解一致。高度硫酸化的GAG，例如肝素，与包含高正电荷密度的互补结合位点硫酸乙酰肝素的肽（或蛋白质内的肽序列）相互作用最紧密，具有更少和更多间隔的负电荷基团，与具有更宽间隔的阳离子残基的肽（或具有蛋白质的肽序列）上的互补位点相互作用最紧密。 (C) 1997 学术出版社
Glycosaminoglycan (GAG)-protein interactions regulate a myriad of physiologic and pathologic processes, yet an understanding of how these molecules interact is lacking, The role of the pattern and spacing of basic amino acids (arginine (R) and lysine (K)) in heparin binding sites was investigated using peptide analogs as well as by examining known heparin binding sites, Peptides having the general structure RnW (n = 3-9, where tyrosine (W) was added for peptide detection) were synthesized and their interaction with heparin was determined by isothermal titration calorimetry, Binding affinity increased with increasing number of R residues, A 9-mer of R (R9W) bound as tightly to heparin as acidic fibroblast growth factor under physiologic conditions, Despite their high affinity for heparin, long stretches of basic amino acids are uncommon in heparin binding proteins, Known heparin binding sites most commonly contain single isolated basic amino acids separated by one nonbasic amino acid, Peptides having the structure, H3CCONH-GRRG(m)RRG(5-m)-CONH2 (denoted as the RRG(m)RR peptide series) and H3CCONH-GRRRG(m)RG(5-m)-CONH2 (denoted as the RRRG(m)R peptide series), where m = 0-5, were synthesized to test the hypothesis that the spacing of basic amino acids in heparin binding sites is optimally arranged to interact with different GAGs, The peptides in both the -RRG(m)RR- and -RRRG(m)R- peptide series, when m = 0, bound most tightly with heparin, as measured by affinity chromatography. In contrast, the -RRG(m)RR-peptide series interacted most tightly with heparan sulfate when m = 0 or 1, whereas the -RRRG(m)R- peptide series bound tightest when m = 3, These results are consistent with our understanding of heparin and heparan sulfate structure, A highly sulfated GAG, such as heparin, interacts most tightly with peptides (or peptide sequences within proteins) containing a complementary binding site of high positive charge density, Heparan sulfate, having fewer and more highly spaced negatively charged groups, interacts most tightly with a complementary site on a peptide (or peptide sequences with proteins) that has more widely spaced cationic residues. (C) 1997 Academic Press