Abstract
A dye displacement method was developed for determination of the affinities of compounds toward the active site of the detergent-solubilized sarcoplasmic reticulum CaATPase. Titration of the enzyme with 2',3'-O-(2,4,6-trinitrophenyl)-ADP resulted in a large absorbance difference in the visible spectrum. Subsequent addition of compounds which bind to the active site causes displacement of the dye; resulting absorbance changes were treated to gain dissociation constants for added ligands. The active site affinities of ATP, and nonhydrolyzable nucleotide analogues were determined. Both occupancy of the high affinity calcium site and added divalent cations have large influences on nucleotide affinity.
Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a callMore From: Biochemical and Biophysical Research Communications
Disclaimer: All third-party content on this website/platform is and will remain the property of their respective owners and is provided on "as is" basis without any warranties, express or implied. Use of third-party content does not indicate any affiliation, sponsorship with or endorsement by them. Any references to third-party content is to identify the corresponding services and shall be considered fair use under The CopyrightLaw.
