Wayne D. Frasch
Wayne D. Frasch joined the faculty at Arizona State University in Fall 1989 where he is now a Professor and a member of the ASU Center for the Study of Early Events in Photosynthesis. He is an active participant in the Biomolecular Nanotechology graduate research training program supported by a National Science Foundation IGERT grant.
Dr. Frasch previously held a faculty position at the University of Michigan in Ann Arbor, and was a visiting research scientist at the Mayo Clinic. He has served on a NIH study section, and has been a Fellow of the Horace Rackham Foundation as well as the Eli Lilly Foundation. In 1985-1986, he served on the NSF/USDA/DOE advisory panel to establish the Plant Science Centers Program. In 1994, he was awarded the Golden Key National Honor Society Award for Excellence in Teaching.
Dr. Fraschs primary research interest concerns the FoF1 ATP synthase. This enzyme is required by virtually every living organism to catalyze the conversion of energy from food or light (during photosynthesis) into ATP, a form of energy that can be used to drive most cellular processes. The enzyme is the smallest molecular motor known. As it operates a core of protein subunits rotates inside a group of stationary subunits. Each of three surrounding stationary subunits contains a catalytic site for the synthesis of ATP from ADP and phosphate that work in an alternating mechanism much like a three cylinder engine.
The F1 portion of the enzyme that contains the catalytic sites can be removed from Fo. Under these conditions the F1 has ATP hydrolysis activity that drives the rotation of the gamma subunit in the inner core of the protein. The rotation of the gamma subunit in single F1 enzyme molecules can be observed under a microscope when the nonrotating subunits are attached to a cover slip and an optical probe is attached to the gamma subunit.
With grant support from the National Institutes of Health, the Frasch laboratory is examining the mechanism with which F1 uses the hydrolysis of ATP to drive the rotation of the gamma subunit. In these studies, site-directed mutagenesis is used to change specific amino acid side chains in the protein. The enzyme that contains each mutant is then evaluated for differences in the ability to hydrolyze ATP and for the ATPase-dependent rotation of the gamma subunit as observed in single molecule experiments. The residues that are receiving the most intense scrutiny have some connection to those that serve as ligands to the magnesium cofactor of the enzyme. In previous work, the Frasch laboratory pioneered the combined use of EPR spectroscopy of vanadyl, a magnesium surrogate for F1, with site-directed mutagenesis to identify how the metal ligands change with each step in catalysis. These ligand changes were shown to be essential to the sequential protein conformational changes in the catalytic mechanism.
Dr. Fraschs laboratory is also using the F1-ATPase as a biomolecular motor in the development of the Molecular Semaphore Device with funding from DARPA. This device uses the ability to detect the rotation of single F1 molecules as a means to detect the hybridization of single molecules of DNA. The device has the potential to increase the sensitivity of detection of DNA microarrays to the ultimate single molecule level, and has received support from the defense department specifically to detect biological warfare agents like Anthrax. Drug testing and proteomics are additional potential applications of this device. The development of the single molecule technology for this device also has direct application to the basic research on the rotational mechanism of the F1-ATPase in the Frasch laboratory.
Dr. Frasch received his Bachelors degree from Hope College in Holland, Michigan and his Ph.D. at the University of Kentucky in Lexington. He was an NSF predoctoral research fellow at the Marine Biological Laboratory, Woods Hole, Massachusetts, and did his postdoctoral work in the Biochemistry Department at the University of Wisconsin in Madison.
Patents
Frasch, W. D., Spetzler, D., and York, J. “High Speed, High Fidelity, High Sensitivity Nucleic Acid Detection”, Provisional Filed, 8/30/06, published 3/6/08; publication number: WO 2008/027977; PCT number: PCT/US2007/0771286.
Chapsky, L., Frasch, W. D., Chou, C., Zenhausern, F., and Goronkin, H. “Single-Molecule Detection of Biological Warfare Agents Using the F1-ATPase Biomolecular Motor” Patent 6,989,235 Awarded, January 2006
Frasch, W. D., Spetzler, D., and York, J. “Methods for Generating a Distribution of Optimal Solutions to Nondeterministic Polynomial Optimization Problems” Provisional filed 1/27/06, serial number 60/762,971.
Frasch, W. D. and Xiong, F. “Padlock-Mediated qRT-PCR to accelerate the Answer Detection Step in DNA Computing. Filed 11/21/06.
Frasch, W. D. and He, Liyan “Single Molecule Detection using Molecular Motors” Provisional patent application filed 12/15/03. International Publication Date: September 1, 2005, International Publication Number: WO 2005/080603 A3
Frasch, W. D. and Chapsky, L.“Polarization-Enhanced Detector with Gold Nanorods for Detecting Nanoscale Rotation” Provisional patent application filed 12/11/02. Nonprovisional application filed 12/10/03. Publication number WO2004/053501 A2
Selected Publications
Spetzler, D., York, J., Martin. J., Ishmukhametov, R., and Frasch, W. D. (2008) "Microsecond Resolution of Enzymatic Movement Using Dark-Field Microscopy", Methods, in press (doi:10.1016/j.ymeth.2008.05.004).
Spetzler, D., Xiong, F., and Frasch, W. D. (2008) "Heuristic solution to a 10-City Traveling Salesman Problem Using Probabilistic DNA Computing", LNCS 4848, 152-160.
York, J., Spetzler, D., Xiong, F., and Frasch, W. D. (2008) "Single Molecule Detection of DNA via Sequence-Specific Links between F1-ATPase Motors and Gold Nanorod Sensors", Lab. Chip 8, 415-419. Among top-10 LOC articles accessed on-line. Highlighted in Chemical Biology, a Royal Society of Chemistry news magazine that provides a snapshot of the latest, most exciting, chemical biology developments.
Hornung, T., Ishmukhametov, R., Spetzler, D., Martin, J., and Frasch, W. D. (2008) "Determination of Torque Generation from the Power Stroke of Escherichia coli F1-ATPase." Biochim. Biophys. Acta- Bioenergetics 1777, 579-582.
Spetzler, D., Xiong, F., and Frasch, W.D. (2007) "Probabilistic DNA Computing Solution to a Fully Connected 10-City Asymmetric Traveling Salesman Problem" Proc. DNA 13, 9-18.
Xiong, F., Spetzler, and Frasch, W. D. (2007) "Elimination of Secondary Structures for DNA Computing", Proc. DNA 13, 241-249.
Spetzler, D., York, J., Dobbin, C., Martin, J., Xiong, F., Ishmukhametov, R., Day, L., Yu, J., Kang, H., Porter, K., Hornung, T., and Frasch, W.D. (2007) "Recent Developments of Biomolecular Motors as On-Chip Devices using Single Molecule Techniques", Lab. Chip 7, 1633-1643. Among top 10 most accessed LOC articles in 2007.
York, J., Spetzler, D., Hornung, T., Ishmukhametov, R., Martin, J., and Frasch, W.D. (2007) "Abundance of Escherichia coli F1-ATPase Molecules Observed to Rotate via Single-Molecule Microscopy with Gold Nanorod Probes", J. Bioenergetics and Biomembranes, 39, 435-439.
Spetzler, D., York, J., Lowry, D., Daniel, D., Fromme, R. and Frasch, W. D. (2006) "Microsecond Time Resolution of Single Molecule F1-ATPase Rotation", Biochemistry 45, 3117-3124. Accelerated publication designated a Hot Article by the ACS because it was among top-10 down loads from the journal for two consecutive months.
Boltz, K. and Frasch, W.D. (2006) "Hydrogen Bonds between the a and b Subunits of the F1-ATPase Allow Communication between the Catalytic site and the Interface of the b Catch Loop and the g Subunit." Biochemistry 45, 11190-11199.
Lowry, D. and Frasch, W. D. (2005) "Interactions between bD372 and g-Subunit N-terminus residues gK9 and gS12 are Important for ATP Synthase Activity Catalyzed by the E. coli FoF1 ATP Synthase" Biochemistry 44, 7275-7281.
Boltz, K. W. and Frasch, W. D. (2005) "Interactions of gT273 and gE275 with the b Subunit PSAV Segment that Links the g-Subunit to the Catalytic Site Walker Homology B Aspartate are Important to the Function of Escherichia coli F1Fo ATP Synthase", Biochemistry 44, 9497-9506.