Click chemistry is a novel, Cu-mediated Hüisgen reaction that is a highly versatile way to make triazoles. In this project, the [3+2] dipolar cycloaddition between azides and terminal alkynes is leveraged to construct N, N bidentate ligands that are likely to be luminescent on their own or upon complexation with metal atoms. |
Ribonuclease inhibitor (RI) is a cytosolic protein that inhibits the activity of pancreatic-type ribonucleases. The RI:ribonuclease complex is the subject of much research because select ribonucleases, for example ribonuclease A (RNase A), show anti-tumor activity when they are able to escape the inhibitory action of RI. Conversely, angiogenin (ANG), a ribonuclease and potent inducer of angiogenesis, may facilitate tumor metastasis by inducing neovascularization. RI may, in the case of ANG, prove a useful chemotherapeutic agent through the inhibition of angiogenesis. Unfortunately, the oxidation-sensitivity of RI renders it difficult to study in laboratory settings and would limit its efficacy as a pharmaceutical agent for preventing angiogenesis. Thus, our research is attempting to create a variant of RI that resists oxidation, but retains its affinity for pancreatic ribonucleases. The instability of RI in oxidative environments arises from its 32 reduced cysteine residues, each of which must remain reduced for proper functioning of the protein. Therefore, we mutated the cysteine residues into amino acids that are not oxidation-sensitive. We divided the cysteines into six groups based on their locations in the primary and tertiary structures of the protein and successfully created six RI variants, each containing a subset of the positions mutated. Analysis of these mutants will allow us to evaluate which, if any, of the positions contribute significantly to the stability of RI or its ability to bind ribonucleases. |
