TOWARDS DETECTING PROTEIN-PROTEIN INTERACTIONS IN SOYBEAN AND OTHER PLANTS VIA MASS SPECTROMETRY
Soybean Genomics and Improvement
2011 Annual Report
1a.Objectives (from AD-416)
Develop methods for targeting and investigating interactions between soybean pathogen proteins and plant proteins involved in disease resistance or other important signaling pathways necessary for optimal plant productivity.
1b.Approach (from AD-416)
Genes from plants and pathogens will be modified to encode a recognizable protein sequence tag. The modified gene will be expressed in plants. The expressed tagged-protein from the transgene will be purified based on tag recognition methodology and approaches will be designed to isolate the tagged protein and it’s potential interacting protein partners. These protein complexes or aggregates will be analyzed by mass spectrometry, which will resolve the individual protein components and thus reveal which proteins are interacting with the tagged protein.
The goal of this research is to develop methods for investigating molecular interactions between proteins, with the intent to apply that knowledge to the parent project goal of seeking resistance mechanisms to pests and pathogens. Genes from plants and pathogens were modified to encode a sequence tag and were expressed in plants. Tagged proteins were isolated from plants to determine if there were any interacting protein partners. Results suggested ethylene has a role in the specific accumulation and interactions between some proteins. The regulation of these proteins is being investigated. Quarterly meetings, annual budget reports, and an annual written progress report are used to monitor project progress.