通过在两个位点引入精氨酸-甘氨酸-天冬氨酸序列构建二价细胞黏附溶菌酶。

Construction of a divalent cell adhesive lysozyme by introducing the Arg-Gly-Asp sequence at two sites.

作者信息

Yamada T, Shimada Y, Uyeda A, Sugiyama S, Kikuchi M

机构信息

Protein Engineering Research Institute, Osaka, Japan.

出版信息

FEBS Lett. 1995 Oct 30;374(2):262-4. doi: 10.1016/0014-5793(95)01123-v.

DOI:10.1016/0014-5793(95)01123-v

PMID:7589549

Abstract

To increase the cell adhesion activity of 74RGD4, an RGDS-inserted mutant between Val74 and Asn75 of human lysozyme, one more site for the RGD introduction was investigated in the lysozyme molecule. We found that 47RGD4 with RGDS in place of AGDR (residues 47 to 50) in a beta-turn region possesses the same level of adhesion activity as that of 74RGD4. The acceptance of the RGD introduction in the beta-turn region of human lysozyme is in good agreement with recent studies on the functional conformation of RGD. We constructed (47,74)RGD4, a mutant containing RGD at two sites, by combining the N-terminal domain of 47RGD4 and the C-terminal domain of 74RGD4. The (47,74)RGD4 lysozyme, with two functional RGD sequences, exhibits even higher cell adhesion activity than that of 74RGD4 or 47RGD4.

摘要

为了提高人溶菌酶74RGD4（一种在人溶菌酶74位缬氨酸和75位天冬酰胺之间插入RGDS的突变体）的细胞黏附活性，研究了溶菌酶分子中另一个可引入RGD的位点。我们发现，在β-转角区域用RGDS取代AGDR（47至50位残基）的47RGD4具有与74RGD4相同水平的黏附活性。人溶菌酶β-转角区域对RGD引入的接受情况与最近关于RGD功能构象的研究结果高度一致。我们通过将47RGD4的N端结构域和74RGD4的C端结构域结合，构建了一个在两个位点含有RGD的突变体(47,74)RGD4。具有两个功能性RGD序列的(47,74)RGD4溶菌酶表现出比74RGD4或47RGD4更高的细胞黏附活性。