From ribonuclease A toward bovine seminal ribonuclease: A step by step thermodynamic analysis

被引:18
作者
Catanzano, F
Graziano, G
Cafaro, V
DAlessio, G
DiDonato, A
Barone, G
机构
[1] UNIV NAPLES FEDERICO II,DIPARTIMENTO CHIM,I-80134 NAPLES,ITALY
[2] UNIV SALERNO,DIPARTIMENTO CHIM,I-84081 BARONISSI,SA,ITALY
[3] UNIV NAPLES FEDERICO II,DIPARTIMENTO CHIM ORGAN & BIOL,I-80134 NAPLES,ITALY
关键词
D O I
10.1021/bi971358j
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
A proline, a leucine, and two cysteine residues, introduced at positions 19, 28, 31, and 32 of bovine pancreatic RNase A, i.e. the positions occupied by these residues in the subunit of bovine seminal RNase, the only dimeric RNase of the pancreatic-type superfamily, transform monomeric RNase A into a dimeric RNase, endowed with the same ability of BS-RNase of swapping its N-terminal segments. The thermodynamic consequences of the progressive introduction of these four residues into RNase A polypeptide chain have been studied by comparing the temperature-and urea-induced denaturation of three mutants of RNase A with that of a stable monomeric derivative of BS-RNase. The denaturation processes proved reversible for all proteins, and well represented by the two-state N double left right arrow D transition model. The progressive introduction of the four residues into RNase A led to a gradual shift of the protein stability toward that characteristic of monomeric BS-RNase, which, in turn, is markedly less stable than RNase A with respect to both temperature-and urea-induced denaturation. On the other hand, the thermal stability of a dimeric active mutant of RNase A is found to approach that of wild-type seminal RNase.
引用
收藏
页码:14403 / 14408
页数:6
相关论文
共 35 条
[1]   THESEUS - A NEW SOFTWARE PACKAGE FOR THE HANDLING AND ANALYSIS OF THERMAL-DENATURATION DATA OF BIOLOGICAL MACROMOLECULES [J].
BARONE, G ;
DELVECCHIO, P ;
FESSAS, D ;
GIANCOLA, C ;
GRAZIANO, G .
JOURNAL OF THERMAL ANALYSIS, 1992, 38 (12) :2779-2790
[2]   THERMAL-DENATURATION OF RIBONUCLEASE-T1 - A DSC STUDY [J].
BARONE, G ;
DELVECCHIO, P ;
FESSAS, D ;
GIANCOLA, C ;
GRAZIANO, G ;
PUCCI, P ;
RICCIO, A ;
RUOPPOLO, M .
JOURNAL OF THERMAL ANALYSIS, 1992, 38 (12) :2791-2802
[3]  
Beintema J.J., 1997, RIBONUCLEASES STRUCT, P245
[4]  
Bevington P. R., 2002, Data Reduction and Error Analysis For the Physical Sciences, V3rd
[5]   Temperature-induced denaturation of ribonuclease S: A thermodynamic study [J].
Catanzano, F ;
Giancola, C ;
Graziano, G ;
Barone, G .
BIOCHEMISTRY, 1996, 35 (41) :13378-13385
[6]   STABILITY OF ALPHA-HELICES [J].
CHAKRABARTTY, A ;
BALDWIN, RL .
ADVANCES IN PROTEIN CHEMISTRY, VOL 46, 1995, 46 :141-176
[7]  
D'Alessio G, 1997, RIBONUCLEASES STRUCT, P383
[8]  
DALESSIO G, 1975, BIOCHEMISTRY-US, V14, P1116, DOI 10.1021/bi00677a004
[9]  
DIDONATO A, 1994, J BIOL CHEM, V269, P17394
[10]   INTERCHAIN DISULFIDE BRIDGES IN RIBONUCLEASE BS-1 [J].
DIDONATO, A ;
DALESSIO, G .
BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS, 1973, 55 (03) :919-928