Streamlined determination of processive run length and mechanochemical coupling of nucleic acid motor activities

Máté Gyimesi, Kata Sarlós, Imre Derényi, Mihály Kovács

Research output: Contribution to journalArticle

10 Citations (Scopus)

Abstract

Quantitative determination of enzymatic rates, processivity and mechanochemical coupling is a key aspect in characterizing nucleotide triphosphate (NTP)-driven nucleic acid motor enzymes, for both basic research and technological applications. Here, we present a streamlined analytical method suitable for the determination of all key functional parameters based on measurement of NTP hydrolysis during interaction of motor enzymes with the nucleic acid track. The proposed method utilizes features of kinetic time courses of NTP hydrolysis that have not been addressed in previous analyses, and also accounts for the effect of protein traps used in kinetic experiments on processivity. This analysis is suitable for rapid and precise assessment of the effects of mutations, physical conditions, binding partners and other effectors on the functioning of translocases, helicases, polymerases and other NTP-consuming processive nucleic acid motors.

Original languageEnglish
Article numbergkq014
Pages (from-to)e102
JournalNucleic acids research
Volume38
Issue number7
DOIs
Publication statusPublished - Jan 29 2010

ASJC Scopus subject areas

  • Genetics

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