Electrostatic interactions across the dimer-dimer interface contribute to the pH-dependent stability of a tetrameric malate dehydrogenase

被引：15

作者：

Bjork, A

Mantzilas, D

Sirevåg, R

Eijsink, VGH

机构：

[1] Univ Oslo, Dept Biol, N-0316 Oslo, Norway

[2] Univ Oslo, Dept Biochem, N-0316 Oslo, Norway

[3] Agr Univ Norway, Dept Chem & Biotechnol, N-1432 As, Norway

来源：

FEBS LETTERS | 2003年 / 553卷 / 03期

关键词：

malate dehydrogenase; oligomerization; thermal stability; electrostatic interactions;

D O I：

10.1016/S0014-5793(03)01076-7

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：

Malate dehydrogenase (MDH) from the moderately thermophilic bacterium Chloroflexus aurantiacus (CaMDH) is a tetrameric enzyme, while MDHs from mesophilic bacteria usually are dimers. Using site-directed mutagenesis, we show here that a network of electrostatic interactions across the extra dimer-dimer interface in CaMDH is important for thermal stability and oligomeric integrity. Stability effects of single point mutations (E25Q, E25K, D56N, D56K) varied from -1.2degreesC to -26.8degreesC, and depended strongly on pH. Gel-filtration experiments indicated that the 26.8degreesC loss in stability observed for the D56K mutant at low pH was accompanied by a shift towards a lower oligomerization state. (C) 2003 Federation of European Biochemical Societies. Published by Elsevier B.V. All rights reserved.

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收藏

页码：423 / 426

页数：4

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