Hope College Department of Physics and Engineering
Research Experiences for Undergraduates
Summer 2006
Project Summary

 

Project Title:

Ion Beam Analysis of Metalloprotein Stoichiometry
Student Name:

Joshua D. Warner (with Lindsay A. Ellsworth)
Student's Home Institution: Hope College
Research Advisor: Drs. Graham F. Peaslee and Paul A. DeYoung
Source of Support: National Science Foundation under Grant Numbers 0354920 and 0319523.

A novel technique for determining the stoichiometric ratio of metal to protein in metalloproteins after polyacrylamide gel electrophoresis is being developed. Native polyacrylamide gel electrophoresis was used for protein separation and provides thin samples necessary for ion beam analysis after drying. Particle induced X-ray emission (PIXE) spectroscopy is a non-destructive ion beam analysis technique excellently suited to determine the concentrations of heavy elements. Particle elastic scattering analysis (PESA) is a similar technique which measures the areal density of a thin target by quantifying the energy loss of the transmitted ions. A combination of PESA and PIXE has been developed to provide a quantitative technique for the determination of stoichiometric ratios in metalloproteins. Initial work has focused on Fe in myoglobin, while current work is focusing on the production of Cu-metallothionein from the CUP1-2 locus in Saccharomyces cerevisiae.

*This material is based upon work supported by the National Science Foundation under NSF-REU Grant No. 0452206

____________________________________________________________________________________________________________________________________________________________

Publications and Presentations:
"Quantitative Analysis of Metalloprotein Stoichiometry with PIXIE and PESA." J.D. Warner*, P.A. DeYoung, L.A. Ellsworth*, M.J. Rycenga*, L.M. Kiessel*, and G.F. Peaslee. Accepted for publication in Nucl. Inst. And Meth. B, (2010).
"PIXE and PESA Analysis of Metalloprotein Stoichiometry." G.F. Peaslee, P.A. DeYoung, L.A. Ellsworth*, L.M. Kiessel*, M.J. Rycenga*, and J.D. Warner*. PIXE 2007, 11th International Conference on Particle-Induced X-Ray Emission and its Analytical Applications. Puebla, Mexico. May 26, 2007.
"Orgin of Finer Grained (<150 micron) Dune Sand Along the Southeastern Coast of Lake Michigan: the Glacial Connection". Eric Johnson, Josh Warner, Graham Peaslee, Paul DeYoung, and Edward Hansen. Annual Meeting of the Geological Society of America, Philadelphia, PA, October 2006. Paper No. 214-11.
"Origin of Finer Grained (¡150 micron) Dune Sand Along the Southeastern Coast of Lake Michigan: the Glacial Connection." Eric Johnson*, Josh Warner*, Graham Peaslee, Paul DeYoung, and Edward Hansen. Paer 214-11. Annual Meeting of the Geological Society of America. Philadelphia PA, October 2006.