130 Preliminary characterization of wheat proteins by HPLC-multi-angle laser light scattering (MALLS).

G. L. Lookhart (1) and S. R. BEAN (2). (1) USDA, ARS, GMPRC, Manhattan, KS 66502; (2) Dept. of Grain Science & Industry, Kansas State Univ., Manhattan, KS 66506.

Multi-angle laser light scattering (MALLS) is capable of providing absolute molecular masses (without the need for generating calibration curves to standards), as well as molecular geometry of biomolecules. MALLS was used in conjunction with HPLC to characterize gliadin and glutenin proteins of wheat. This methodology has previously been applied to the study of wheat starch by several research groups. Here we describe preliminary studies on the use of this technique for the study of gluten proteins. Reversed phase (RP)-HPLC-MALLS is being applied to study individual glutenin and gliadin proteins. Preliminary data gathered on 5 HMW-GS show that the masses determined by RP-HPLC-MALLS closely matched those of their respective cDNA masses (<5%). Unreduced polymeric proteins are being characterized with size exclusion chromatography (SEC)-MALLS. Development of extraction conditions (solvents, stirring/vortexing, temperature, sonication, etc.) for analysis of unreduced proteins by SEC-MALLS is in progress. The use of HPLC-MALLS provides an easy, automated, high resolution method for further characterizing both reduced and unreduced glutenin proteins.