Skip to main content
Article thumbnail
Location of Repository

Probing nucleotide dissociation from myosin in vitro using microgram quantities of myosin

By Richard J. Clark, Miklos Nyitrai, Martin Webber and Michael A. Geeves

Abstract

The detailed kinetic analysis of novel myosin motors is often limited by the quantity of stable protein available for study. We show here that the use of coumarin based fluorescent ADP analogues allows the assay of ADP affinities and dissociation rate constants in a flash photolysis apparatus using microg quantities of the rabbit muscle myosin S1. We go on to use the analogues to characterise two other rat muscle myosin S1 and the motor domain of Dictyostelium cytoplasmic myosin II. The results show that the fluorescence change for the binding of a coumarin based ADP analogue to a myosin motor domain is variable in sign as well as amplitude for the different proteins. The analysis also provided estimates of the affinities of caged-ATP for S1 which were < or = 10 microM for muscle S1s and > 200 microM for the non-muscle myosin

Topics: Q
Publisher: Springer
Year: 2003
OAI identifier: oai:kar.kent.ac.uk:13192
Download PDF:
Sorry, we are unable to provide the full text but you may find it at the following location(s):
  • http://dx.doi.org/10.1023/A:10... (external link)
  • Suggested articles


    To submit an update or takedown request for this paper, please submit an Update/Correction/Removal Request.