| The FOLPROCOM consortium is united through their commom interests in utilizing viral systems to tackle problems of protein folding, production and macromolecular assembly. We propose an interdisciplinary, multipronged attack that will focus on the following issues: To improve the yield of soluble protein, assays will be developed for the rapid detection and characterization of folding and aggregation inside cells and in vitro. These will then be used to tailor the best media, host/vector/chaperone combination for expression of different viral proteins. Chaperone-assisted folding will be explored by identifying novel chaperones of the endoplasmic reticulum and exploring their potential for increasing secretion with model proteins. The use of chaperones in in vitro folding from inclusion bodies will also be explored. An alternative to these methods will be to target proteins to the reoviral viroplasm, an intracellular factory about which little is known, but is readily isolatable. The production of fibrous proteins, retroviral Gag polyprotein and macromolecular complexes, e.g. the retroviral pre-integration complex and dsRNA virus cores, will be key areas where new expression and isolation methods will be tested for their effectiveness. The acquired knowledge and technologies will improve expression of pathogenic virus proteins for vaccine and anti-viral drug development. |
