FRET IN ALPHA-1-ANTITRYPSIN:CHIMOTRYPSIN COMPLEXES

这个子项目是许多研究子项目中的一个 由NIH/NCRR资助的中心赠款提供的资源。子项目和 研究者（PI）可能从另一个NIH来源获得了主要资金， 因此可以在其他CRISP条目中表示。所列机构为 研究中心，而研究中心不一定是研究者所在的机构。 对α-1-抗胰蛋白酶的三种突变体进行工程改造，使它们可以用荧光体作为荧光供体进行标记。然后通过加入蛋白酶胰凝乳蛋白酶来形成复合物，胰凝乳蛋白酶本身用受体荧光体标记。共振能量转移预期发生后，激发的供体发色团在抗胰蛋白酶，其程度取决于该蛋白质复合物的特定结构和距离。使用荧光衰减测量TCSPC，本项目的目标是阐明抗胰蛋白酶，丝氨酸蛋白酶抑制剂，和蛋白酶糜蛋白酶之间的复合物的结构和动力学信息。

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.