Protein anatomy: structure and function of peptide fragments corresponding to the secondary structure units of barnase.
Globular proteins can be decomposed into several modules or secondary structure units. It is useful to investigate the functions of such structural units in order to understand the folding units of proteins. In our previous work, barnase was divided into six peptide fragments corresponding to modules, and some of them were shown to have RNA-binding and RNase activity [Yanagawa, et al. (1993) J. Biol. Chem. 268, 5861-5865]. Barnase mutant proteins obtained by permutation of the structural units also had RNase activity [Tsuji, T. et al. (1999) J. Mol. Biol. 286, 1581-1596]. Here we investigated the structure and function of peptide fragments corresponding to secondary structure units of barnase. The results of circular dichroism spectroscopy indicated that some of the peptide fragments form helical structures in aqueous solutions containing over 30% 2,2,2-trifluoroethanol, and the S6 (94-110) peptide fragment is induced to form a beta-sheet structure in the presence of RNA. The S6 peptide fragment forms aggregate complexes with RNA. Electron microscopic analysis showed that the aggregate complexes were comprised of filaments. These results indicate that not only modules but also secondary structure units dissected from a globular protein have functional and structure-forming capabilities.
Ishizaka
M
M
Department of Biology, Kyushu University, Hakozaki, Higashi-ku, Fukuoka 812-8581, Japan. hyana@applc.keio.ac.jp
Ohshima
Y
Y
Yanagawa
H
H
D000595:Amino Acid Sequence; D001426:Bacterial Proteins; D002942:Circular Dichroism; D006863:Hydrogen-Ion Concentration; D008854:Microscopy, Electron; D008958:Models, Molecular; D008969:Molecular Sequence Data; D010446:Peptide Fragments / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism* / Q000648:ultrastructure; D017510:Protein Folding; D017433:Protein Structure, Secondary / Q000187:drug effects; D017434:Protein Structure, Tertiary / Q000187:drug effects; D012341:RNA, Ribosomal, 5S / Q000378:metabolism / Q000494:pharmacology / Q000648:ultrastructure; D016601:RNA-Binding Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D012260:Ribonucleases / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism* / Q000648:ultrastructure; D017384:Sequence Deletion / Q000235:genetics; D012996:Solutions; D013329:Structure-Activity Relationship; D013696:Temperature; D014270:Trifluoroethanol
D001426:Bacterial Proteins; D010446:Peptide Fragments; D012341:RNA, Ribosomal, 5S; D016601:RNA-Binding Proteins; D012996:Solutions; D014270:Trifluoroethanol; D012260:Ribonucleases; C019760:Bacillus amyloliquefaciens ribonuclease