Structural and functional consequences of substitutions at the Pro108-Arg14 hydrogen bond in bovine adrenodoxin

被引:12
作者
Grinberg, A [1 ]
Bernhardt, R [1 ]
机构
[1] Univ Saarland, Fachbereich Pharm & Umwelttechnol, Fachrichtung Biochem 12 4, D-66041 Saarbrucken, Germany
关键词
D O I
10.1006/bbrc.1998.9225
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
Elimination of Pro108 in bovine adrenodoxin is known to result in the formation of a misfolded protein that is not able to incorporate a [2Fe-2S] cluster and rapidly degrades upon expression in E. coli, However, no experimental explanation for this phenomenon has been demonstrated so far. Using the recently obtained 3D structure of the truncated mutant Adx(4-108) we have studied the reasons of the protein stabilization by the proline residue by means of site-directed mutagenesis, Two main results have been obtained: (i) the conserved hydrogen bond Pro108-Arg14, that connects different structural domains of Adx, contributes 6 kJ/mol into the protein stability and (ii) the presence of proline at position 108 provides a low conformational entropy of the unfolded state, supporting a gain in the Gibbs energy of 5.4 kJ/mol at 37 degrees C. (C) 1998 Academic Press.
引用
收藏
页码:933 / 937
页数:5
相关论文
共 13 条
[1]  
AKHREM AA, 1979, ACTA BIOL MED GER, V38, P257
[2]  
BECKERT V, 1994, J BIOL CHEM, V269, P2568
[3]   Specific aspects of electron transfer from adrenodoxin to cytochromes P450scc and P45011 beta [J].
Beckert, V ;
Bernhardt, R .
JOURNAL OF BIOLOGICAL CHEMISTRY, 1997, 272 (08) :4883-4888
[4]   Cytochrome P450: Structure, function, and generation of reactive oxygen species [J].
Bernhardt, R .
REVIEWS OF PHYSIOLOGY BIOCHEMISTRY AND PHARMACOLOGY, VOL 127, 1996, 127 :137-221
[5]  
BUROVA TV, 1995, PROTEIN SCI, V4, P909
[6]   HYDROGEN-BONDING AND BIOLOGICAL SPECIFICITY ANALYZED BY PROTEIN ENGINEERING [J].
FERSHT, AR ;
SHI, JP ;
KNILLJONES, J ;
LOWE, DM ;
WILKINSON, AJ ;
BLOW, DM ;
BRICK, P ;
CARTER, P ;
WAYE, MMY ;
WINTER, G .
NATURE, 1985, 314 (6008) :235-238
[7]  
Kimura T., 1968, STRUCTURE BONDING, V5, P1
[8]   Energetics of protein structure [J].
Makhatadze, GI ;
Privalov, PL .
ADVANCES IN PROTEIN CHEMISTRY, VOL 47, 1995, 47 :307-425
[9]   New aspects of electron transfer revealed by the crystal structure of a truncated bovine adrenodoxin, Adx(4-108) [J].
Müller, A ;
Müller, JJ ;
Muller, YA ;
Uhlmann, H ;
Bernhardt, R ;
Heinemann, U .
STRUCTURE, 1998, 6 (03) :269-280
[10]   Forces contributing to the conformational stability of proteins [J].
Pace, CN ;
Shirley, BA ;
McNutt, M ;
Gajiwala, K .
FASEB JOURNAL, 1996, 10 (01) :75-83