PMID:2171503
| Citation |
Kanaya, S, Kimura, S, Katsuda, C and Ikehara, M (1990) Role of cysteine residues in ribonuclease H from Escherichia coli. Site-directed mutagenesis and chemical modification. Biochem. J. 271:59-66 |
|---|---|
| Abstract |
The role of the three cysteine residues at positions 13, 63 and 133 in Escherichia coli RNAase H, an enzyme that is sensitive to N-ethylmaleimide [Berkower, Leis & Hurwitz (1973) J. Biol. Chem. 248, 5914-5921], was examined by using both site-directed mutagenesis and chemical modification. Novel aspects that were found are as follows. First, none of the cysteine residues is required for activity. Secondly, chemical modification of either Cys-13 or Cys-133 with thiol-blocking reagents inactivates the enzyme, but that of Cys-63 does not. Thus the sensitivity of E. coli RNAase H to N-ethylmaleimide arises not from blocking of the thiol group but from steric hindrance by the modifying group incorporated at either Cys-13 or Cys-133. |
| Links | |
| Keywords |
Amino Acid Sequence; Chemical Phenomena; Chemistry; Chromatography, High Pressure Liquid; Circular Dichroism; Cysteine; Dithionitrobenzoic Acid; Endoribonucleases/antagonists & inhibitors; Endoribonucleases/chemistry; Endoribonucleases/metabolism; Escherichia coli/enzymology; Ethylmaleimide/pharmacology; Iodoacetates/pharmacology; Iodoacetic Acid; Molecular Sequence Data; Mutagenesis, Site-Directed; Peptide Mapping; Ribonuclease H; Serine Endopeptidases/metabolism; Structure-Activity Relationship |
| edit table |
Significance
You can help EcoliWiki by summarizing why this paper is useful
Useful Materials and Methods
You can help Ecoliwiki by describing the useful materials (strains, plasmids, antibodies, etc) described in this paper.
Annotations
<protect><annotationlinks/></protect>
EcoliWiki Links
Add links to pages that link here (e.g. gene, product, method pages)
See also
References
See Help:References for how to manage references in EcoliWiki.
