Origin of dimeric structure in the ribonuclease superfamily

被引:55
作者
Ciglic, MI
Jackson, PJ
Raillard, SA
Haugg, M
Jermann, TM
Opitz, JG
Trabesinger-Rüf, N
Benner, SA [1 ]
机构
[1] Univ Florida, Dept Chem, Div Biochem, Gainesville, FL 32611 USA
[2] Univ Florida, Dept Anat & Cell Biol, Gainesville, FL 32611 USA
[3] Perkin Elmer, Appl Biosyst Div, Warrington WA3 7PB, Cheshire, England
关键词
D O I
10.1021/bi972203e
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 enable application of postgenomic evolutionary approaches to understand the divergence of behavior and function in ribonucleases (RNases), the impact of divergent sequence on the divergence of tertiary and quaternary structure is analyzed in bovine pancreatic and seminal ribonucleases, which differ by 23 amino acids, In a crystal, seminal RNase is a homodimer joined by two "antiparallel" intersubunit disulfide bonds between Cys-31 from one subunit and Cys-32' from the other and having composite active sites arising from the "swap" of residues 1-20 from each subunit. Specialized Edman degradation techniques have completed the structural characterization of the dimer hi solution, new crosslinking methods have been developed to assess the swap, and sequence determinants of quaternary structure have been explored by protein engineering using the reconstructed evolutionary history of the protein family as a guide. A single Cys at either position 32 (the first to be introduced during the divergent evolution of the family) or 31 converts monomeric RNase A into a dimer. Even with an additional Phe at position 31, another residue introduced early in the seminal lineage, swap is minimal, A hydrophobic contact formed by Leu-28, however, also introduced early in the seminal lineage, increases the amount of "antiparallel" connectivity of the two subunits and facilitates swapping of residues 1-20. Efficient swapping requires addition of a Pro at position 19, a residue also introduced early in the divergent evolution of the seminal RNase gene. Additional cysteines required for dimer formation are found to slow refolding of the protein through formation of incorrect disulfide bonds, suggesting a paradox in the biosynthesis of the protein. Further studies showed that the dimeric form of seminal RNase known in the crystal is not the only form in vivo, where a substantial amount of heterodimer is known, These data complete the acquisition of the background needed to understand the evolution of new structure, behavior, and function in the seminal RNase family of proteins.
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页码:4008 / 4022
页数:15
相关论文
共 97 条
[71]   REACQUISITION OF QUATERNARY STRUCTURE BY FULLY REDUCED AND DENATURED SEMINAL RIBONUCLEASE [J].
PARENTE, A ;
DALESSIO, G .
EUROPEAN JOURNAL OF BIOCHEMISTRY, 1985, 149 (02) :381-387
[72]   THE DUAL-MODE QUATERNARY STRUCTURE OF SEMINAL RNASE [J].
PICCOLI, R ;
TAMBURRINI, M ;
PICCIALLI, G ;
DIDONATO, A ;
PARENTE, A ;
DALESSIO, G .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1992, 89 (05) :1870-1874
[73]   THE DESIGN OF SYNTHETIC GENES [J].
PRESNELL, SR ;
BENNER, SA .
NUCLEIC ACIDS RESEARCH, 1988, 16 (05) :1693-1702
[74]   Selective removal of ribonucleases from solution with covalently anchored macromolecular inhibitor [J].
Rahman, MH ;
Kang, I ;
Waterbury, RG ;
Narang, U ;
Bright, FV ;
Wang, JH .
ANALYTICAL CHEMISTRY, 1996, 68 (01) :134-138
[75]  
RAILLARD SA, 1993, THESIS SWISS FEDERAL
[76]   LOCALIZATION OF 2 INTERCHAIN DISULFIDE BRIDGES IN DIMERS OF BOVINE ALPHA-S2-CASEIN - PARALLEL AND ANTIPARALLEL ALIGNMENTS OF THE POLYPEPTIDE-CHAINS [J].
RASMUSSEN, LK ;
HOJRUP, P ;
PETERSEN, TE .
EUROPEAN JOURNAL OF BIOCHEMISTRY, 1992, 203 (03) :381-386
[77]   NEW PYRIMIDINE-SPECIFIC RIBONUCLEASE FROM BOVINE SEMINAL PLASMA THAT IS ACTIVE ON BOTH SINGLE AND DOUBLE-STRANDED POLYRIBONUCLEOTIDES AND THAT CAN DISTINGUISH BETWEEN MG2+-CONTAINING AND MG2+-DEPLETED NATURALLY-OCCURRING RNAS [J].
REDDY, ESP ;
SITARAM, N ;
BHARGAVA, PM ;
SCHEIT, KH .
JOURNAL OF MOLECULAR BIOLOGY, 1979, 135 (03) :525-544
[78]  
Richards, 1971, ENZYMES, P647
[79]  
RICHARDS FM, 1959, J BIOL CHEM, V234, P1459
[80]   MOLECULAR-CLONING OF THE HUMAN EOSINOPHIL-DERIVED NEUROTOXIN - A MEMBER OF THE RIBONUCLEASE GENE FAMILY [J].
ROSENBERG, HF ;
TENEN, DG ;
ACKERMAN, SJ .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1989, 86 (12) :4460-4464