If
protein is Homo-tetramer then one can expect the identical binding sites. I am also working
on homo-dimeric protein which binds to DNA. I used PRISM to estimate the binding affinity through
flourescence binding method using “SATURATION and
NON-LINEAR REGRESSION and ONE SITE binding Model’ considering protein
concentration as dimer or monomer. Then
I looked for the sensibility of the model by analyzing (comparing) best fit
values (like SD, Bmax) of the parameters with reasonable certainty.
Unlike ITC binding model fitting (gives good estimate of stoichiometry, cooperativity
and also binding sites (one, two or sequential binding sites)), flourescence binding models do not give very good estimate of
these parameters. Thus, based on protein
you should assume the model which fits to the properties of the protein.
Good luck
Raj
Dear All,Firstly sorry for asking a non-crystallography question, but i want help in understanding the data analysis for fitting a protein-ligand binding data.Actually i have a protein which is a tetramer in solution and i have done its flourescence binding with a ligand. I am trying to fit the data to a 4-site binding model in scientist. But i donot have a correct model to fit in the data for identical or non-identical, co-operative or sequential binding. Can anyone help me in analysing the binding data.Any help will be highly appreciated.Thankyou !