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ARS Home » Pacific West Area » Albany, California » Western Regional Research Center » Produce Safety and Microbiology Research » Research » Research Project #426378

Research Project: Identifying Host Proteins that Facilitate the Assembly of Viral and Phage Capsids and Small Molecules that Interfere with this Assembly

Location: Produce Safety and Microbiology Research

Project Number: 2030-42000-051-02-T
Project Type: Trust Fund Cooperative Agreement

Start Date: Apr 1, 2014
End Date: Mar 31, 2019

Objective:
The goal of this project is to identify proteins involved in facilitating the assembly of viral capsids. These proteins will be identified by their binding to small molecule-based affinity resins. These small molecules have been previously demonstrated to interfere with viral capsid assembly. This will facilitate the identification of proteins which bind to small molecules known to interfere with viral capsid assembly.

Approach:
Proprietary small molecules will be covalently attached to an immobilized resin to produce an affinity resin. Extracts from cultivated cells or tissues containing desirable protein fractions will be passed over the affinity resin. Those proteins that bind to the affinity resin will be eluted from the resin with progressively more stringent washes. The set of washes will prepared to run on an SDS-PAGE gel. The gel will be stained with Coomassie blue. The observed protein bands will be excised. The excised bands will be robotically digested and then analyzed by mass spectrometry. The mass spectrometry-based analysis will be used to identify the proteins that bound to the affinity resin.