Hope College Physics Department
Research Experiences for Undergraduates
Summer 2012
Project Summary

 

Project Title: Stoichiometric Analysis of Metalloproteins in Thin Polyacrylamide Gels by Simultaneous Particle-Induced X-ray Emission Spectroscopy and Nuclear Reaction Analysis
Student Name: Megan Sibley
Student's Home Institution: Clemson University
Research Advisor: Dr. Paul DeYoung and Dr. Graham Peaslee
Source of Support:

This work is supported by the National Science Foundation under NSF-REU Grant No. PHY/DMR-1004811.

Particle-Induced X-ray Emission Spectroscopy (PIXE) and Nuclear Reaction Analysis (NRA) are two highly sensitive ion beam analytical techniques which probe the elemental constitution and areal density, respectively, of a sample. Using a 1.7 MV tandem electrostatic particle accelerator, these methods have been utilized simultaneously to determine the identity and stoichiometry of metal atoms within protein samples of known metal content; results indicate the technique is a useful analytical tool which can be accurately extended to proteins of hitherto unknown metal stoichiometries. The nondestructive technique, originally developed for dried protein spots on a suitably thin support, has been extended for metalloproteins suspended in polyacrylamide gel. This allows use of the technique in biochemical applications in which proteins are separated by polyacrylamide gel electrophoresis (PAGE), after which proteins are not easily removed from the gel. Various methods of sample preparation such as sample mounting and gel drying have been considered. Additionally, such factors as the thickness, constitution, and possible contaminants of the gel have been investigated and are reported.

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