Document Detail


Study of RNase A mechanism and folding by means of synthetic 63-residue analogs.
MedLine Citation:
PMID:  833149     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
A 63-residue RNase A analog containing residues 26 to 35 then alanine, 41 to 59 and 73 to 84 then glycine, 100 to 110 then glycine, and 117 to 124 was synthesized by the solid phase method. The deletions comprised ordered (an alpha helix, parts of the beta sheet) and less ordered structures including 27 of the 56 residues invariant in mammalian ribonucleases. The synthetic 63-residue analog was cleaved from the insoluble support with liquid HF, reduced-reoxidized, fractionated by gel filtration, and purified further on an affinity column specific for the active site fold of RNase A. It had an activity of 8 to 14 per cent in the transphosphorylation step using poly(C) and poly(U) as substrates. It also had low synthetic and hydrolytic activity (0.2 per cent) and showed RNase A-like specificity toward the substrates tested. This indicated that all residues essential for substrate binding and catalysis were present and that their relative positions in the three-dimensional structure were probably very similar to those in native RNase A. Therefore, structure-function studies with the 63-residue RNase A analog should allow conclusions about the mode of action of the natural enzyme. As a first step in this direction, lysine 41 which is believed to be important for catalysis was replaced in the 63-residue analog by tyrosine or glutamine. The resulting (Tyr-41)- and (Gln 41)-63-residue analogs were also bound by the affinity column and had the same substrate specificity as native RNase A. They differed from each other, from the (Lys 41)-63-residue analog, and the 124-residue natural enzyme only with respect to the relative rates of the catalyzed reactions. Thus, lysine 41 does not seem to be essential for the functioning of RNase A.
Authors:
B Gutte
Related Documents :
10092459 - Crystal structures of phenylalanyl-trna synthetase complexed with phenylalanine and a p...
1841379 - Structure-function relationship of hammerhead ribozymes as probed by 2'-modifications.
10882069 - Capture and visualization of a catalytic rna enzyme-product complex using crystal latti...
10673029 - Characterization of the azoarcus ribozyme: tight binding to guanosine and substrate by ...
15956979 - The catalytic diversity of rnas.
22618019 - Fluorescent probe for detection of bacteria: conformational trigger upon bacterial redu...
22362299 - Protease inhibitor monotherapy: what is its role?
670189 - Characterization of phosphate oxygen exchange reactions catalyzed by myosin through mea...
11279119 - Adjacent basic amino acid residues recognized by the cop i complex and ubiquitination g...
Publication Detail:
Type:  Journal Article    
Journal Detail:
Title:  The Journal of biological chemistry     Volume:  252     ISSN:  0021-9258     ISO Abbreviation:  J. Biol. Chem.     Publication Date:  1977 Jan 
Date Detail:
Created Date:  1977-03-15     Completed Date:  1977-03-15     Revised Date:  2000-12-18    
Medline Journal Info:
Nlm Unique ID:  2985121R     Medline TA:  J Biol Chem     Country:  UNITED STATES    
Other Details:
Languages:  eng     Pagination:  663-70     Citation Subset:  IM    
Export Citation:
APA/MLA Format     Download EndNote     Download BibTex
MeSH Terms
Descriptor/Qualifier:
Amino Acid Sequence
Amino Acids / analysis
Binding Sites
Models, Molecular
Peptide Fragments / chemical synthesis,  metabolism
Protein Binding
Protein Conformation
Ribonucleases* / chemical synthesis,  metabolism
Chemical
Reg. No./Substance:
0/Amino Acids; 0/Peptide Fragments; EC 3.1.-/Ribonucleases

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine


Previous Document:  Mechanism of induction of 3-hydroxy-3-methylglutaryl coenzyme A reductase in human leukocytes.
Next Document:  Topography of the external surface of the human red blood cell membrane studied with a nonpenetratin...