Interaction of heparin with a synthetic pentadecapeptide from the C-terminal heparin-binding domain of fibronectin.
Interaction of heparin with a synthetic pentadecapeptide from the C-terminal heparin-binding domain of fibronectin.
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肝素与纤连蛋白 C 端肝素结合域的合成十五肽的相互作用。
DOI：
10.1021/bi9926734
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发表时间：
2000
期刊：
影响因子：
2.9
通讯作者：
Rabenstein,DL
中科院分区：
文献类型：
--
作者：
Hari,SP;McAllister,H;Chuang,WL;Christ,MD;Rabenstein,DL
合成的十五肽FN-C/H II（KNNQKSEPLIGRKKT-NH 2）具有纤维连接蛋白模块III 14的羧基末端肝素结合结构域的序列。用~ 1H、~（23）Na NMR和肝素亲和层析研究了FN-C/H Ⅱ与牛肺肝素的相互作用。FN-C/H II从pD <2至pD> 10与肝素结合;在较高pD下，随着赖氨酸侧链铵基团的滴定，结合降低。Na+抗衡离子被FN-C/H II从围绕肝素钠的抗衡离子冷凝体积中置换，这提供了结合涉及静电相互作用的直接证据。FN-C/H II的五个铵基中的每一个的pKa值在与肝素结合时增加，这与化学位移数据一起表明结合涉及FN-C/H II的铵基与肝素的羧酸根和/或硫酸根基团之间的离域和直接静电相互作用。NMR数据也提供了证据的精氨酸侧链的胍基团与肝素上的阴离子位点的直接相互作用。肝素对FN-C/H II和13个类似肽的亲和力随着碱性残基数目的增加而增加，其中赖氨酸和精氨酸残基被丙氨酸系统地取代。各赖氨酸和精氨酸对肝素对FN-C/H II的亲和力的相对贡献为R12> K13> K14> K1> K5。核奥弗豪泽增强（NOE）数据表明，虽然FN-C/H II在水溶液中很大程度上是非结构化的，但结合的肽在肽的L9− T15片段上的重叠，转弯状构象之间相互转化。FN-C/H II与肝素衍生的六糖相互作用的NOE数据，以及通过23 Na NMR测定的FN-C/H II从肝素中置换的Na+离子的数量，表明肽与肝素的六糖片段结合。FN-C/H II和itsd-氨基酸类似物肽与肝素的相互作用获得了相同的NMR和肝素亲和色谱结果，这对于肽作为其中细胞粘附起关键作用的疾病的治疗剂的潜在用途是有意义的。
The synthetic pentadecapeptide FN-C/H II (KNNQKSEPLIGRKKT-NH2) has the sequence of the carboxy-terminal heparin-binding domain of module III14of fibronectin. Interaction of FN-C/H II with bovine lung heparin has been studied by1H and23Na NMR spectroscopy and by heparin affinity chromatography. FN-C/H II binds to heparin from pD <2 up to pD ∼10; at higher pD, the binding decreases as the lysine side-chain ammonium groups are titrated. Na+counterions are displaced from the counterion condensation volume that surrounds sodium heparinate by FN-C/H II, which provides direct evidence that the binding involves electrostatic interactions. The pKAvalues for each of the five ammonium groups of FN-C/H II increase upon binding to heparin which, together with chemical shift data, indicates that the binding involves both delocalized and direct electrostatic interactions between ammonium groups of FN-C/H II and carboxylate and/or sulfate groups of heparin. NMR data also provide evidence for the direct interaction of the guanidinium group of the arginine side chain with anionic sites on heparin. The affinity of heparin for FN-C/H II and for 13 analogue peptides in which lysine and arginine residues were systematically substituted with alanine increases as the number of basic residues increases. The relative contribution of each lysine and arginine to the affinity of heparin for FN-C/H II is R12> K13> K14> K1> K5. Nuclear Overhauser enhancement (NOE) data indicate that, while FN-C/H II is largely unstructured in aqueous solution, the bound peptide interconverts among overlapping, turn-like conformations over the L9− T15segment of the peptide. NOE data for the interaction of FN-C/H II with a heparin-derived hexasaccharide, together with the number of Na+ions displaced from heparin by FN-C/H II as determined by23Na NMR, indicates that the peptide binds to a hexasaccharide segment of heparin. Identical NMR and heparin affinity chromatography results were obtained for the interaction of FN-C/H II and itsd-amino acid analogue peptide with heparin, which is of interest for the potential use of peptides as therapeutic agents for diseases in which cell adhesion plays a critical role.