CAVITY MUTANTS OF SAVINASE(TM) - CRYSTAL-STRUCTURES AND DIFFERENTIAL SCANNING CALORIMETRY EXPERIMENTS GIVE HINTS OF THE FUNCTION OF THE BURIED WATER-MOLECULES IN SUBTILISINS

被引:22
作者
PEDERSEN, JT
OLSEN, OH
BETZEL, C
ESCHENBURG, S
BRANNER, S
HASTRUP, S
机构
[1] NOVO NORDISK AS,DK-2880 BAGSVAERD,DENMARK
[2] CTR ADV RES BIOTECHNOL,ROCKVILLE,MD 20850
[3] DESY,EUROPEAN MOLEC BIOL LAB,D-22603 HAMBURG,GERMANY
关键词
SAVINASE(TM); SUBTILISIN; BURIED WATER; MUTANTS; X-RAY STRUCTURES;
D O I
10.1006/jmbi.1994.1572
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
The subtilisin molecule possesses several internal water molecules, which may be characterised as an integral part of the protein structure. We have introduced specific mutations (T7lI, T7lS, T71V, T71A and T71C) at position 71 in the subtilisin variant Savinase(TM) from Bacillus lentus. This position is involved in a hydrogen bonded network with several internal water molecules, forming a water channel. The water channel and most of the other internal water molecules are positioned in the interface between two half-domains of the subtilisin molecule. The data presented here indicate that the internal water molecules are structural, and may be the result of trapping during the folding process.
引用
收藏
页码:193 / 202
页数:10
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