This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. The approach has been developed to determine the pKas of the titratable residues by two-dimensional NMR. The pKa determination on titratable residues has the indispensable advangtage of understanding the interresidual interaction involved in the single protein and also the interaction between the proteinase and their inhibitors. In our search, OMTKY3 variants work as a good model for proteinase inhibitor. So far,we have finished the pH titration for different OMTKY3 variants. And more than 400 two-dimensional TOCSY spectra have been successfully obtained. Based upon the pKa result we got, the interaction between OMTKY3 variants and chymotrypsin have been characterized by the enzyme-substrate model.Also, some intramolecular through-space interaction(H bond, etc.) have also been identified.
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