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Recently, an unexpected, rapid exponential ATP\u2010stimulated DNA unwinding rate was observed from an <jats:italic>Archaeoglobus fulgidus<\/jats:italic> helicase (AfXPB) as compared to the slower conventional helicases from <jats:italic>Sulfolobus tokodaii<\/jats:italic>, StXPB1 and StXPB2. This unusual rapid activity suggests a \u201cmolecular wrench\u201d mechanism arising from the torque applied by AfXPB on the duplex structure in transitioning from open to closed conformations. However, much remains to be understood. Here, we investigate the concentration dependence of DNA helicase binding and ATP\u2010stimulated kinetics of StXPB2 and AfXPB, as well as their binding and activity in Bax1 complexes, via an electrochemical assay with redox\u2010active DNA monolayers. StXPB2 ATP\u2010stimulated activity is concentration\u2010independent from 8 to 200\u2009nM. Unexpectedly, AfXPB activity is concentration\u2010dependent in this range, with exponential rate constants varying from seconds at concentrations greater than 20\u2009nM to thousands of seconds at lower concentrations. At 20\u2009nM, rapid exponential signal decay ensues, linearly reverses, and resumes with a slower exponential decay. This change in AfXPB activity as a function of its concentration is rationalized as the crossover between the fast molecular wrench and slower conventional helicase modes. AfXPB\u2010Bax1 inhibits rapid activity, whereas the StXPB2\u2010Bax1 complex induces rapid kinetics at higher concentrations. This activity is rationalized with the crystal structures of these complexes. These findings illuminate the different physical models governing molecular wrench activity for improved biological insight into a key factor in DNA repair.<\/jats:p>","DOI":"10.1002\/pro.4815","type":"journal-article","created":{"date-parts":[[2023,10,24]],"date-time":"2023-10-24T14:20:23Z","timestamp":1698157223000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":6,"title":["Molecular wrench activity of <scp>DNA<\/scp> helicases: Keys to modulation of rapid kinetics in <scp>DNA<\/scp> repair"],"prefix":"10.1002","volume":"32","author":[{"given":"Ashan P.","family":"Wettasinghe","sequence":"first","affiliation":[{"name":"Department of Physics The University of Texas at Dallas  Richardson Texas USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Melodee O.","family":"Seifi","sequence":"additional","affiliation":[{"name":"Department of Physics The University of Texas at Dallas  Richardson Texas 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