UBIQUITIN CHAIN ANALYSIS

摘要

There is described a method for analyzing ubiquitin polymers using linkage-specific deubiquitinase enzymes. Novel specificities of deubiquitinase enzymes are also provided.

主权项

1. A method for determining the linkage type in a polyubiquitin chain, comprising the steps of:
(a) contacting an ubiquitin polymer with a first linkage-specific deubiquitinase under conditions in which the enzyme catalyses the disassociation of one or more, but less than seven, ubiquitin linkages; (b) analysing the product of the catalysis for the presence of lower molecular weight fractions of the ubiquitin polymer; and (c) repeating steps (a) and (b) in the presence of a second, different linkage-specific deubiquitinase.