Current objectives include methods to characterize the products of the reaction of nitroxide spin traps with enzymatically generated free methods to assay for protein phosphorylation without the requirement improved assays for phospholipase C, and improved methods for the study of lipid peroxidation. An analytical high pressure liquid chromatographic method For the separation of POBN- linolate radical adducts has been developed. The fractions are characterized by their UV and FT-IR spectra, with attempts to obtain mass spectra in progress. A convenient assay for phospholipase C has been developed that permits the comparative use of a variety of substrates with low milliunit sensitivity. We are currently working on an approach to the assay of protein kinase C that will involve the use of pre-labeled protein substrates. A recommended approach to the use of scopoletin for measurement of trace levels of hydrogen peroxide gives much higher reproducibility than earlier methods.
