Nuclease having altered specificity

Details Nuclease having altered specificity Nuclease having altered specificity

1994-01-21

1995-02-14

Baker, Keith

Chemistry: molecular biology and microbiology

Enzyme , proenzyme; compositions thereof; process for...

Hydrolase

435195, 435810, C12N 922

Patent

active

053895371

ABSTRACT:
A ribonuclease molecule altered at a single amino acid, relative to its wild-type form, displays altered substrate specificity and substrate binding mechanism. The altered protein cleaves RNA efficiently after C, U and A residues, whereas the wild-type protein cannot cleave efficiently after A. The change that alters the specificity also permits the protein to cleave poly (A) portions of an RNA molecule processively.

REFERENCES:
Beintema, J. J., "Structure, Properties and Molecular Evolution of Pancreatic-Type Ribonucleases," Life Chemistry Reports, 4:333-389 (1987).
Deutscher, M. P., "The Metabolic Role of RNases," TIBS, 13:136-139 (1988).
"New Products for Molecular Biology," Boehringer Mannheim Catalog, p. 8 (Jul. 1993).
Ribonuclease Protection Assay information pamphlet, Ambion, 1993.
Guardian.TM. RNase Protection Assay Kit, CLONTECHniques, Clontech, Apr. 1993.
Meador III, J. et al., "Purification and characterization of Escherichia coli RNase I: Comparisons with RNase M," Eur. J. Biochem., 198:549-553 (1990).
RNase One.TM., Promega Catalog, p. 59 (1992-1993).
Raines, R. T., "Engineering to Explore and Exploit Ribonuclease A Function, " (Abstract), presented at Scientific meeting in Capri, Italy, May 11, 1993.
Ehretsmann, C. P. et al., "mRNA degradation in procaryotes," FASEB Journal6:3186-3192 (1992).
Gutti (1978) J. Biol. Chem. 263, 3837-3842.

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