Cross-linked aggregates of multimeric enzymes:: A simple and efficient methodology to stabilize their quaternary structure

被引：85

作者：

Wilson, L ^{[1
]}

Betancor, L ^{[1
]}

Fernández-Lorente, G ^{[1
]}

Fuentes, M ^{[1
]}

Hidalgo, A ^{[1
]}

Guisàn, JM ^{[1
]}

Pessela, BCC ^{[1
]}

Fernández-Lafuente, R ^{[1
]}

机构：

[1] CSIC, Inst Catalysis, Dept Biocatalysts, E-28049 Madrid, Spain

来源：

BIOMACROMOLECULES | 2004年 / 5卷 / 03期

关键词：

D O I：

10.1021/bm034528i

中图分类号：

Q5 [生物化学]; Q7 [分子生物学];

学科分类号：

071010 ; 081704 ;

摘要：

In this manuscript, we show that the immobilization of proteins following the technique of cross-linked protein aggregates (CLEAS) may permit the stabilization of the most complex multimeric enzymes by preventing their dissociation. To illustrate that, we have first prepared CLEAS with two tetrameric catalases. Activity recovery was over 40%, and no protein subunit could be desorbed from the CLEAS after boiling in SDS. More interestingly, the enzyme stability, which in its soluble form strongly depends on the enzyme concentration, becomes fully independent of this parameter. This permitted the enzyme stability to greatly increase under diluted conditions. In fact, diluted CLEAs presented a higher stability than those of their glyoxyl derivatives counterparts, which were unable to fully stabilize the multimeric structure of these tetrameric enzymes.

引用

页码：814 / 817

页数：4