Method for in vitro inactivation of RNase S

Details Method for in vitro inactivation of RNase S Method for in vitro inactivation of RNase S

1996-04-29

1998-10-06

Gitomer, Ralph

Chemistry: molecular biology and microbiology

Measuring or testing process involving enzymes or...

435 23, 435 24, C12Q 100

Patent

active

058174559

ABSTRACT:
A method for inactivating, at a desired stage of an in vitro process, a target enzyme having coupled thereto a biotin molecule, includes adding to a reaction mix an inactivating protein having an affinity for the biotin molecule that is sufficient to inhibit the activity of the target enzyme.
The method is embodied in the ribonuclease activity of the enzyme RNase S, which can be active in a form composed of an S peptide and an S protein, not covalently bound together, which associate to form the catalytic molecule. By adding an affinity moeity to the S peptide, it is possible to a second, inactivating protein having affinity for the affinity moeity to disassociate the S peptide from the S protein, and thereby terminate catalytic activity of RNase S at a desired point in any reaction.

REFERENCES:
patent: 4745054 (1988-05-01), Rabin
patent: 5028524 (1991-07-01), Fujisawa
patent: 5057412 (1991-10-01), Rabin
patent: 5563033 (1996-10-01), Lawrence et al.

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