CAREER: Measurement, modeling and control of molecular motors above a nanopore

University Of California-Santa Cruz, Santa Cruz CA

Investigators

Abstract

Intellectual Merit: Novel approaches for single molecule measurement and manipulation are required to uncover with high resolution the dynamics and function of biological macromolecules. The proposed research will optimize a biological nanopore instrument for measuring and controlling individual molecular motors at sub-millisecond resolution. In preliminary work, active voltage control regulates a single DNA in a nanopore for repeated binding and dissociation of individual enzymes above the nanopore. The first research aim is to reduce the total latency of detecting DNA-enzyme complex formation. Modeling will then be employed to estimate equilibrium and binding rate constants from the measured DNA-enzyme complex assembly data. The final aim is measuring and controlling individual lambda exonculease motors during degradation of duplex DNA into single-stranded DNA above the nanopore. Active control and modeling will permit exploration of voltage dependence in pausing and in the translocation step in the catalytic cycle of lambda exonuclease on the nanopore, at the single molecule level. Broader Impacts: The primary goal of the education plan is to advance feedback control as the basis for interdisciplinary outreach efforts, in the form of high school, undergraduate and graduate courses at UCSC. The PI has co-designed an introductory course on dynamics and control, which is taught in summer to high school students, and each fall and spring to freshmen and sophomores at UCSC. A graduate course on the role of feedback control in single molecule instrumentation, cross-listed for computer and electrical engineering and bioengineering students, will also be developed to disseminate the more advanced topics of the research plan.