Rate-limiting Pyrophosphate Release by HIV Reverse Transcriptase Improves Fidelity.

ABSTRACT: Previous measurements of the rates of polymerization and pyrophosphate release with DNA templates showed that pyrophosphate (PP_i) dissociation was fast after nucleotide incorporation so that it did not contribute to enzyme specificity (k_cat/K_m). Here, kinetic parameters governing nucleotide incorporation and PP_i release were determined using an RNA template. Compared with a DNA template of the same sequence, the rate of chemistry increased by up to 10-fold (250 versus 24 s^-1), whereas the rate of PP_i release decreased to approximately 58 s^-1 so that PP_i release became the rate-limiting step. During processive nucleotide incorporation, the first nucleotide (TTP) was incorporated at a fast rate (152 s

SUBMITTER: Li A 

PROVIDER: S-EPMC5159514 | biostudies-literature | 2016 Dec

REPOSITORIES: biostudies-literature