Identification of the critical residues responsible for differential reactivation of the triosephosphate isomerases of two trypanosomes.

ABSTRACT: The reactivation of triosephosphate isomerase (TIM) from unfolded monomers induced by guanidine hydrochloride involves different amino acids of its sequence in different stages of protein refolding. We describe a systematic mutagenesis method to find critical residues for certain physico-chemical properties of a protein. The two similar TIMs of Trypanosoma brucei and Trypanosoma cruzi have different reactivation velocities and efficiencies. We used a small number of chimeric enzymes, additive mutants and planned site-directed mutants to produce an enzyme from T. brucei with 13 mutations in its sequence, which reactivates fast and efficiently like wild-type (WT) TIM from T. cruzi, and another enzyme from T. cruzi, with 13 slightly altered mutations, which reactivated slowly and inefficientl

SUBMITTER: Rodriguez-Bolanos M 

PROVIDER: S-EPMC5090059 | biostudies-literature | 2016 Oct

REPOSITORIES: biostudies-literature