Simple and universal method to determine dissociation constants for enzyme/ligand complexes.
A simple and in principle universal method is proposed for measuring enzyme/ligand dissociation constants. The method is based on measuring enzyme activity remaining after heat treatment in the absence and in the presence of ligands. The method is especially suitable for enzymes interacting with nucleosides, nucleosides and oligonucleotides since for such enzymes convenient spectrophotometric assays are available.
Bzowska
Agnieszka
A
Department of Biophysics, Institute of Experimental Physics, University of Warsaw, Zwirki i Wigury 93, 02-089 Warsaw, Poland. abzowska@biogeo.uw.edu.pl
Magnowska
Lucyna
L
D000225:Adenine / Q000031:analogs & derivatives / Q000737:chemistry; D001671:Biochemistry / Q000379:methods; D004795:Enzyme Stability; D006147:Guanine / Q000737:chemistry; D006358:Hot Temperature; D007700:Kinetics; D008024:Ligands; D009943:Organophosphorus Compounds / Q000737:chemistry; D010710:Phosphates / Q000737:chemistry; D011683:Purine-Nucleoside Phosphorylase / Q000032:analysis* / Q000737:chemistry; D013053:Spectrophotometry
C090125:9-(2-phosphonylmethoxypropyl)-2,6-diaminopurine; D008024:Ligands; D009943:Organophosphorus Compounds; D010710:Phosphates; D006147:Guanine; D011683:Purine-Nucleoside Phosphorylase; D000225:Adenine