[16] Measurement of protein dissociation constants by tritium exchange

摘要: Publisher Summary This chapter presents a radioisotope method for determining protein dissociation constants, that circumvents the difficulties encountered in conventional kinetic experiments, or interposed by alteration—that is, trace amounts of tritium and carbon-14 are incorporated into protein. Because isotope trace, retains its natural properties. The is exchanged with solvent hydrogen at rates sensitive to state (folded, unfolded, associated, dissociated), which range over several orders magnitude so as be very much less than form interconversion readily measurable wide concentrations, including dilute solutions. remains affixed protein, thereby providing radioactivity, measure concentration, even nanomolar concentrations.