X-ray crystallographic studies of RNase A variants engineered at the most destabilizing positions of the main hydrophobic core: Further insight into protein stability

被引:7
作者
Kurpiewska, Katarzyna [1 ]
Font, Josep [2 ]
Ribo, Marc [2 ]
Vilanova, Maria [2 ]
Lewinski, Krzysztof [1 ]
机构
[1] Jagiellonian Univ, Fac Chem, PL-30060 Krakow, Poland
[2] Univ Girona, Lab Engn Prot, Dept Biol, Fac Ciencies, Girona 17071, Spain
关键词
crystal structure; ribonuclease A; mutation-structure relationship; cavity; stability; CAVITY-CREATING MUTATIONS; RIBONUCLEASE-A; CRYSTAL-STRUCTURES; PACKING INTERACTIONS; HYDROGEN-EXCHANGE; ACTIVE-SITE; PRESSURE; TEMPERATURE; RESIDUES; MUTANTS;
D O I
10.1002/prot.22480
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
To investigate the structural origin of decreased pressure and temperature stability, the crystal structure of bovine pancreatic ribonuclease A variants V47A, V54A, V57A, 181A, 1106A, and V108A was solved at 1.4-2.0 angstrom resolution and compared with the structure of wild-type protein. The introduced mutations had only minor influence on the global structure of ribonuclease A. The structural changes had individual character that depends on the localization of mutated residue, however, they seemed to expand from mutation site to the rest of the structure. Several different parameters have been evaluated to find correlation with decrease of free energy of unfolding Delta Delta G(T), and the most significant correlation was found for main cavity volume change. Analysis of the difference distance matrices revealed that the ribonuclease A molecule is organized into five relatively rigid subdomains with individual response to mutation. This behavior consistent with results of unfolding experiments is an intrinsic feature of ribonuclease A that might be surviving remnants of folding intermediates and reflects the dynamic nature of the molecule. Proteins 2009; 77:658-669. (C) 2009 Wiley-Liss, Inc.
引用
收藏
页码:658 / 669
页数:12
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