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 tool which can be accurately extended to proteins of hitherto unknown metal stoichiometry.  The nondestructive technique, originally developed for dried protein spots on a 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).  Various methods of sample preparation and gel drying have been considered.  Additionally, such factors as the thickness, constitution, and possible contaminants of the gel have been investigated.

Stoichiometric analysis of metalloproteins by simultaneous particle-induced X-ray emission spectroscopy and nuclear reaction analysis

245th National Meeting (2013)

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