Binding of benzamidine and protons to trypsin as measured by difference spectra☆

Add time:09/08/2019    Source:sciencedirect.com

The interaction of trypsin and benzamidine causes changes in their ultraviolet absorption spectra; the spectral changes are measured by difference spectroscopy, and it appears that both components contribute to the difference spectrum. The dependence of the height of a difference peak at 248 mμ upon the concentrations of benzamidine and hydrogen ion fits a model in which the two ligands compete for a single class of non-interacting sites on trypsin. The system is characterized by the dissociation constants Kl = 2 × 10−5m and Kh = 2.4 × 10−5m, for benzamidine and protons, respectively. A similar difference spectrum is obtained with diisopropyl phosphoryltrypsin, whereas none is obtained with p-toluene sulfonyl-l-lysine chloro ketonetrypsin or with trypsinogen.

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