title: A procedure for analysis of stopped-flow transients for protein-ligand association.
abstract: A method for extracting kinetic and optical parameters from progress curves for protein-ligand association, obtained by stopped-flow experiments, is described. The method is limited to one-step and two-step association kinetics, but it allows concentration of protein and offset of the signals to be adjustable parameters during an interactive non-linear least-squares fitting procedure. The method is tested on simulated pseudo-experimental data and applied to progress curves obtained in a stopped-flow spectrofluorimeter, for association of the translation initiation factor eIF4E with 7-methyl-GDP, an analog of 5'-end of mRNA.
authors.affiliation: Department of Biophysics, Warsaw University, Warswa 02-089, Poland.
authors.forename: M
authors.initials: M
authors.lastname: Długosz
authors.forename: E
authors.initials: E
authors.lastname: Bojarska
authors.forename: J M
authors.initials: JM
authors.lastname: Antosiewicz
mesh_terms: D000465:Algorithms; D001671:Biochemistry / Q000379:methods*; D004305:Dose-Response Relationship, Drug; D039561:Eukaryotic Initiation Factor-4E; D006153:Guanosine Diphosphate / Q000031:analogs & derivatives* / Q000737:chemistry / Q000378:metabolism; D007700:Kinetics; D008024:Ligands; D015233:Models, Statistical; D010448:Peptide Initiation Factors / Q000737:chemistry*; D011485:Protein Binding; D012333:RNA, Messenger / Q000378:metabolism; D013050:Spectrometry, Fluorescence / Q000379:methods*
chemical_list: D039561:Eukaryotic Initiation Factor-4E; D008024:Ligands; D010448:Peptide Initiation Factors; D012333:RNA, Messenger; C111106:7-methylguanosine 5'-diphosphate; D006153:Guanosine Diphosphate