TY  - JOUR
T1  - Interrogating the activities of conformational deformed enzyme by single-molecule fluorescence-magnetic tweezers microscopy
LA  - en
PB  - National Academy of Sciences
YR  - 2015
UL  - http://buscaintegrada.pucsp.br/vufind/Record/pubmed-4653173
AB  - Characterizing the impact of fluctuating enzyme conformation on enzymatic activity is critical in understanding the structure–function relationship and enzymatic reaction dynamics. Different from studying enzyme conformations under a denaturing condition, it is highly informative to manipulate the conformation of an enzyme under an enzymatic reaction condition while monitoring the real-time enzymatic activity changes simultaneously. By perturbing conformation of horseradish peroxidase (HRP) molecules using our home-developed single-molecule total internal reflection magnetic tweezers, we successfully manipulated the enzymatic conformation and probed the enzymatic activity changes of HRP in a catalyzed H(2)O(2–)amplex red reaction. We also observed a significant tolerance of the enzyme activity to the enzyme conformational perturbation. Our results provide a further understanding of the relation between enzyme behavior and enzymatic conformational fluctuation, enzyme–substrate interactions, enzyme–substrate active complex formation, and protein folding–binding interactions.
KW  - Biological Sciences
ER  -