METHOD AND SYSTEMS FOR INVESTIGATING THE UBIQUITIN-PROTEASOME SYSTEM

摘要

A method is provided for investigating molecular interactions and enzyme activities related to the ubiquitin-proteasome system (UPS), the method comprising: a) providing at least one surface with a target moiety bound to at least a portion thereof; b) i) exposing said surface to a medium comprising a labelled ubiquitin moiety such as ubiquitin, SUMO, NEDD8 e.t.c., and a component or components of the ubiquitination system suitable to conjugate the ubiquitin moiety to a target moiety; and/or ii) wherein said target moiety is conjugated to a labelled ubiquitin moiety exposing said at least one surface to a medium comprising a deubiquitinating enzyme (DUB) suitable to remove the ubiquitin moiety from a target moiety; c) providing conditions permissive for the conjugation/removal of the labelled ubiquitin moiety to/from the target moiety; and d) detecting conjugation and/or removal of the labelled ubiquitin moiety by detecting labelled ubiquitin moiety located at or near the surface.