Valine 108, a chain-folding initiation site-belonging residue, crucial for the ribonuclease a stability

被引:18
作者
Coll, MG
Protasevich, II
Torrent, J
Ribó, M
Lobachov, VM
Makarov, AA
Vilanova, M [1 ]
机构
[1] Univ Girona, Lab Engn Prot, Dept Biol, Fac Ciencies, Girona 17071, Spain
[2] Russian Acad Sci, VA Engelhardt Mol Biol Inst, Moscow 117984, Russia
基金
俄罗斯基础研究基金会;
关键词
D O I
10.1006/bbrc.1999.1672
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
Thermal denaturation of bovine pancreatic ribonuclease A and a set of its single variants, carrying replacements of hydrophobic residues in the postulated 106-118 chain folding initiation site, has been studied by differential scanning calorimetry. Ribonuclease A variants undergo a two-state thermal transition denaturation except for those with replacement of valine 108. Most mutations cause a significant destabilization of the protein compared to the wild-type, thus demonstrating the importance of hydrophobic residues at the 106-118 region in maintaining the stability of the molecule. Among them, those of valine 108 promote the greatest (14-27 degrees C) destabilization of the molecule. Therefore, valine 108 plays a crucial role for ribonuclease A stability. (C) 1999 Academic Press.
引用
收藏
页码:356 / 360
页数:5
相关论文
共 29 条
[1]   PROTEIN STABILITY CURVES [J].
BECKTEL, WJ ;
SCHELLMAN, JA .
BIOPOLYMERS, 1987, 26 (11) :1859-1877
[2]  
CUCHILLO CM, 1997, RIBONUCLEASES STRUCT, P271
[3]   ENGINEERING RIBONUCLEASE-A - PRODUCTION, PURIFICATION AND CHARACTERIZATION OF WILD-TYPE ENZYME AND MUTANTS AT GLN11 [J].
DELCARDAYRE, SB ;
RIBO, M ;
YOKEL, EM ;
QUIRK, DJ ;
RUTTER, WJ ;
RAINES, RT .
PROTEIN ENGINEERING, 1995, 8 (03) :261-273
[4]   Folding and unfolding kinetics of the proline-to-alanine mutants of bovine pancreatic ribonuclease A [J].
Dodge, RW ;
Scheraga, HA .
BIOCHEMISTRY, 1996, 35 (05) :1548-1559
[5]   CALCULATION OF PROTEIN EXTINCTION COEFFICIENTS FROM AMINO-ACID SEQUENCE DATA [J].
GILL, SC ;
VONHIPPEL, PH .
ANALYTICAL BIOCHEMISTRY, 1989, 182 (02) :319-326
[6]   A specific transition state for S-peptide combining with folded S-protein and then refolding [J].
Goldberg, JM ;
Baldwin, RL .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1999, 96 (05) :2019-2024
[7]   VOLUME CHANGES ON PROTEIN-FOLDING [J].
HARPAZ, Y ;
GERSTEIN, M ;
CHOTHIA, C .
STRUCTURE, 1994, 2 (07) :641-649
[8]   INFORMATION-CONTENT IN THE CIRCULAR-DICHROISM OF PROTEINS [J].
HENNESSEY, JP ;
JOHNSON, WC .
BIOCHEMISTRY, 1981, 20 (05) :1085-1094
[9]   Structure of a hydrophobically collapsed intermediate on the conformational folding pathway of ribonuclease A probed by hydrogen-deuterium exchange [J].
Houry, WA ;
Scheraga, HA .
BIOCHEMISTRY, 1996, 35 (36) :11734-11746
[10]  
JUNCOSAGINESTA M, 1994, BIOTECHNIQUES, V16, P3437