Immobilized amines and basic amino acids as mimetic heparin-binding domains for cell surface proteoglycan-mediated adhesion.
Immobilized amines and basic amino acids as mimetic heparin-binding domains for cell surface proteoglycan-mediated adhesion.
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固定胺和碱性氨基酸作为模拟肝素结合域，用于细胞表面蛋白聚糖介导的粘附。
DOI：
10.1016/s0021-9258(19)50210-1
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发表时间：
1992
期刊：
影响因子：
--
通讯作者：
Jeffrey A. Hubbell
中科院分区：
文献类型：
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作者：
S. Massia;Jeffrey A. Hubbell
二胺共价偶联到玻璃基板促进人包皮成纤维细胞粘附在血清的情况下。这些二胺衍生的基板通过偶联乙二胺，N-甲基氨基乙胺，和N，N-二甲基氨基乙胺（NNDMAEA），磺酰氯活化的玻璃。化学分析的电子光谱表明，二胺通过其伯胺末端偶联，以产生通过两个碳间隔基连接到游离氨基部分的表面结合的仲胺。含有游离叔胺的NNDMAEA修饰底物支持最高程度的细胞铺展（73 +/- 7%活跃铺展细胞）和最广泛的细胞骨架组织。游离的叔胺和表面结合的仲胺都被证明是细胞铺展所需的。赖氨酸和甘氨酸接枝基板支持细胞扩散和细胞骨架组织类似的NNDMAEA修饰的基板上。虽然在这些基质上的铺展细胞内观察到一些应力纤维，但没有形成焦点接触。肝素酶处理不会抑制细胞在二胺衍生底物上的附着或铺展，但软骨素酶ABC抑制细胞在所有底物上的附着和铺展;肝素酶在较小程度上抑制了在赖氨酸和精氨酸衍生底物上的铺展。这些结果表明，这些基板的细胞附着介导的主要是由细胞表面硫酸软骨素蛋白聚糖。这项研究表明，共价接枝NNDMAEA，赖氨酸和精氨酸可以模拟细胞粘附蛋白的糖胺聚糖结合结构域的粘附促进活性。这项研究还表明，这些蛋白聚糖的相互作用取决于在一个非常敏感的方式上的特定结构的固定化胺。
Diamines covalently coupled to glass substrates promoted human foreskin fibroblast adhesion in the absence of serum. These diamine-derivatized substrates were produced by coupling ethylene diamine, N-methylaminoethylamine, and N,N-dimethylaminoethylamine (NNDMAEA), to sulfonyl chloride-activated glass. Electron spectroscopy for chemical analysis demonstrated that the diamines were coupled via their primary amine ends to produce a surface-bound secondary amine linked to a free amino moiety via a two-carbon spacer. NNDMAEA-modified substrates containing free tertiary amines supported the highest degree of cell spreading (73 +/- 7% actively spreading cells) and the most extensive cytoskeletal organization. Both the free tertiary and surface-bound secondary amines were shown to be required for cell spreading. Lysine- and arginine-grafted substrates supported cell spreading and cytoskeletal organization similar to that on NNDMAEA-modified substrates. Although some stress fibers were observed within spread cells on these substrates, focal contacts did not form. Heparinase treatment did not inhibit cell attachment or spreading to the diamine-derivatized substrates, however chondroitinase ABC inhibited cell attachment and spreading on all substrates; heparinase inhibited spreading on lysine- and arginine-derivatized substrates to a lesser extent. These results imply that cell attachment to these substrates was mediated primarily by cell surface chondroitin sulfate proteoglycans. This study demonstrates that covalently grafted NNDMAEA, lysine, and arginine can mimic the adhesion-promoting activity of the glycosaminoglycan-binding domains of cell adhesion proteins. This study also demonstrates that the interaction with these proteoglycans depends in a very sensitive manner on the particular structure of the immobilized amine.