I'm doimg some enzymatic kinetic studies and I need your help!!I'm using a FRET based assay to evaluate the strenght of some potential inhibitors against HIV-1 protease.
I wanted to measure the Km and the Vmax of HIV-1 protease for my peptidic substrate http://www.sigmaaldrich.com/catalog/product/sigma/h0790?lang=it®ion=IT and I performed different enzymatic reaction at 8 substrate concentrations. I used graphpad to obtain Km and Vmax, but the Vmax is expressed as RFU/sec and I know I should convert it as mol/min/mg of protein. How can I do this?
I think I should have a calibration curve of my product but I'm not sure...
thanks a lot