Engineering carboxypeptidase G2 circular permutations for the design of an autoinhibited enzyme.

ABSTRACT: Carboxypeptidase G2 (CPG2) is an Food and Drug Administration (FDA)-approved enzyme drug used to treat methotrexate (MTX) toxicity in cancer patients receiving MTX treatment. It has also been used in directed enzyme-prodrug chemotherapy, but this strategy has been hampered by off-site activation of the prodrug by the circulating enzyme. The development of a tumor protease activatable CPG2, which could be achieved using a circular permutation of CPG2 fused to an inactivating 'prodomain', would aid in these applications. We report the development of a protease accessibility-based screen to identify candidate sites for circular permutation in proximity of the CPG2 active site. The resulting six circular permutants showed similar expression, structure, thermal stability, and, in four cases, ac

SUBMITTER: Yachnin BJ 

PROVIDER: S-EPMC6283397 | biostudies-literature | 2017 Apr

REPOSITORIES: biostudies-literature