Chemical conjugation of linear and cyclic RGD moieties to a recombinant elastin-mimetic polypeptide - A versatile approach towards bioactive protein hydrogels
Chemical conjugation of linear and cyclic RGD moieties to a recombinant elastin-mimetic polypeptide - A versatile approach towards bioactive protein hydrogels
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DOI：
10.1002/mabi.200700234
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发表时间：
2008-06-11
影响因子：
4.6
通讯作者：
Weberskirch, Ralf
中科院分区：
文献类型：
--
作者：
Kaufmann, Doris;Fiedler, Anne;Weberskirch, Ralf
具有氨基酸序列GRDPSS [VPGVG VPGKG VPGVG VPGVG VPEG VPGIG]（7）的弹性蛋白模拟多肽（EMM）（7）用于各种整联蛋白配体（RGD肽）的化学缀合以制备生物活性水凝胶。化学方法涉及（1）用Fmoc或Boc基团化学保护赖氨酸残基，（2）将具有或不具有己酸间隔基的受保护的线性或环状R ⑶配体化学连接至谷氨酸残基，（3）赖氨酸官能团和R ⑶部分的脱保护和（4）交联以形成生物活性水凝胶。1H-1 NMR光谱用于定量反应中的多个步骤。发现Fmoc和Boc的化学保护分别在65%和93%之间。研究的配体包括线性RGD细胞结合[H-FGRGDS-OH（1- 1-RGD）、H-Ahx-FGRGDS-OH（2-Ahx-FGRGDS）和环状-H2 N-（CH 2）（6）COHN-环（-RGDfK-）（H-Ahx-c（-RGDfK-））肽，也具有己酸间隔基。细胞与小鼠成骨细胞的粘附依赖于配体类型、配体密度和间隔物的使用。
An elastin-mimetic polypeptide, (EMM)(7), with the amino-acid sequence GRDPSS [VPGVG VPGKG VPGVG VPGVG VPGEG VPGIG](7) was used for chemical conjugation of various integrin ligands (RGD peptides) to prepare bioactive hydrogels. The chemical approach involved (1) chemical protection of lysine residues with Fmoc or Boc groups, (2) chemical ligation of a protected linear or cyclic RGD ligand, with or without a hexanoic-acid spacer to the glutamic acid residue, (3) deprotection of the lysine functionalities and the RGD moieties and (4) cross-linking to form a bioactive hydrogel. H-1 NMR spectroscopy was used to quantify the multiple steps in the reaction. The chemical protection was found to be between 65 and 93% for Fmoc and Boc, respectively. The ligands studied included linear RGD cell-binding [H-FGRGDS-OH (1-l-RGD), H-Ahx-FGRGDS-OH (2-Ahx-FGRGDS) and a cyclic -H2N-(CH2)(6)COHN-cyclo(-RGDfK-) (H-Ahx-c(-RGDfK-)) peptide also with a hexanoic-acid spacer. Cell adhesion with mouse osteoblast cells was dependent on the ligand type, ligand density and the use of a spacer.