2007 Annual Report 1a.Objectives (from AD-416) To characterize proteins from the brush border membrane (BBM) of Tribolium castaneum to identify novel targets for insect control. 1b.Approach (from AD-416) A database of predicted proteins will be obtained from the annotated T. castaneum genome project, which will greatly facilitate the identification of unknown proteins. Protein from the BBM of larvae will be isolated using standard biochemical techniques to optimize protein recovery from membranes. Samples will be fractionated, and purified proteins will be identified using MALDI-TOF and MS/MS and algorithms for peptide mass fingerprinting. A database will be constructed of the subproteome of BBM proteins, and proteins will be selected for future studies, in particular, those that are critical to digestion and microbial defensive response. 3.Progress Report This report serves to document research conducted under a specific cooperative agreement, between ARS and Kansas State University. Additional details of the research can be found in the report for the parent CRIS 5430-43000-026-00D, Genomics and Proteomics of Stored-Product Insects for Development of New Biopesticides. This research is directly related to Objective 1, to develop proteomic technologies to study proteins in the coleopteran gut that interact with inhibitors and toxins. New control products are needed for beetle storage pests, because the current products are being lost due to insecticide resistance and deregulation. In the second year of a collaboration with Kansas State University Biotechnology Core Facility, we are using proteomics to identify targets for the development of insect control products. ARS personnel have been trained in using proteomics equipment, and two Kansas State University students working in the KSU Core Facility are working on projects to identify proteins in the gut of coloeopteran storage pests. This information will be used to identify beetle midgut proteins that have potential as targets for new insect control products. Monitoring activities include direct supervision of the ARS personnel and technical guidance of the KSU students, monthly meetings to discuss research results, and plan future experiments.