FRET IN ALPHA-1-ANTITRYPSIN:CHIMOTRYPSIN COMPLEXES

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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. Three mutants of alpha-1-antitrypsin were engineered so that they could be labeled with a fluorophor acting as fluorescence donor. Complex formation is then achieved by addition of the proteinase chymptrypsin, which is itself labeled with an acceptor fluorophor. Resonance energy transfer is expected to take place after excitation of the donor chromophor in antitrypsin, its extent depending on the specific structure and distance of this protein complex. Using fluorescence decay measurements by TCSPC, the goal of this project is to elucidate structural and kinetic information of the complex between antitrypsin, a serpin, and the protease chymotrypsin.