Functional assembly of a randomly cleaved protein.
Open Access
- 1 March 1992
- journal article
- research article
- Published by Proceedings of the National Academy of Sciences in Proceedings of the National Academy of Sciences
- Vol. 89 (5) , 1880-1884
- https://doi.org/10.1073/pnas.89.5.1880
Abstract
The sequence of a 939-amino acid polypeptide that is a member of the aminoacyl-tRNA synthetase class of enzymes has been aligned with sequences of 15 related proteins. This alignment guided the design of 18 fragment pairs that were tested for internal sequence complementarity by reconstitution of enzyme activity. Reconstitution was achieved with fragments that divide the protein at both nonconserved and conserved sequences, including locations proximal to or within elements believed to form critical elements of secondary structure. Structure assembly is sufficiently flexible to accommodate fusion of short segments of unrelated sequences at fragment junctions. Complementary chain packing interactions and chain flexibility appear to be widely distributed throughout the sequence and are sufficient to reconstruct large three-dimensional structures from an array of disconnected pieces. The results may have implications for the evolution and assembly of large proteins.Keywords
This publication has 53 references indexed in Scilit:
- Lysine 335, part of the KMSKS signature sequence, plays a crucial role in the amino acid activation catalysed by the methionyl-tRNA synthetase from Escherichia coliJournal of Molecular Biology, 1991
- Assembly of a class I tRNA synthetase from products of an artificially split geneBiochemistry, 1991
- Crystallographic study at 2·5 Å resolution of the interaction of methionyl-tRNA synthetase from Escherichia coli with ATPJournal of Molecular Biology, 1990
- Identification of the tRNA anticodon recognition site of Escherichia coli methionyl-tRNA synthetaseBiochemistry, 1990
- Evidence from Cassette Mutagenesis for a Structure-Function Motif in a Protein of Unknown StructureScience, 1988
- The valyl-tRNA synthetase from Bacillus stearothermophilus has considerable sequence homology with the isoleucyl-tRNA synthetase from Escherichia coliBiochemistry, 1987
- Structure of the Yeast Isoleucyl-tRNA Synthetase Gene (ILS1). DNA-Sequence, Amino-Acid Sequence of Proteolytic Peptides of the Enzyme and Comparison of the Structure to those of other Known Aminoacyl-tRNA SynthetasesBiological Chemistry Hoppe-Seyler, 1987
- Methionyl-tRNA synthetase from Escherichia coli: primary structure at the binding site for the 3'-end of tRNAfMetBiochemistry, 1985
- The nature of the accessible and buried surfaces in proteinsJournal of Molecular Biology, 1976
- A general method applicable to the search for similarities in the amino acid sequence of two proteinsJournal of Molecular Biology, 1970