Abstract

Specific Aims 1. Express HTLV-III encoded wild-type and mutant proteins in E. coli or eukaryotic cells using recombinant DNA clones provided by Drs. Haseltine and Sodrowski at the Dana Farber Cancer Institute (DFCI). This may require additional modification of the encoding DNA sequences. 2. Prepare large scale cultures (both bacterial and eukaryotic) expression proteins of interest. 3. Purify large amounts of expressed proteins in a biologically active form. Renaturation of purified proteins to an active configuration may be required. 4. Provide expressed protein preparations to scientists from DFCI for testing of biological activity. 5. Provide expressed protein preparations to scientists from Agouron Pharmaceuticals, Inc. for crystallography.

  Funding Agency

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Program Projects (P01)
Project #
5P01GM039599-03
Application #
3096344
Study Section
(SRC)
Project Start
1987-09-01
Project End
1992-08-31
Budget Start
1989-09-01
Budget End
1990-08-31
Support Year
3
Fiscal Year
1989
Total Cost
Indirect Cost

  Institution

Name
Agouron Pharmaceuticals, Inc.
Department
Type
DUNS #
City
San Diego
State
CA
Country
United States
Zip Code
92121

  Publications

Goldman, B B; Wipke, W T (2000) QSD quadratic shape descriptors. 2. Molecular docking using quadratic shape descriptors (QSDock). Proteins 38:79-94
Goldman, B B; Wipke, W T (2000) Quadratic shape descriptors. 1. Rapid superposition of dissimilar molecules using geometrically invariant surface descriptors. J Chem Inf Comput Sci 40:644-58
Hostomsky, Z; Hudson, G O; Rahmati, S et al. (1992) RNase D, a reported new activity associated with HIV-1 reverse transcriptase, displays the same cleavage specificity as Escherichia coli RNase III. Nucleic Acids Res 20:5819-24
Hostomsky, Z; Hostomska, Z; Hudson, G O et al. (1991) Reconstitution in vitro of RNase H activity by using purified N-terminal and C-terminal domains of human immunodeficiency virus type 1 reverse transcriptase. Proc Natl Acad Sci U S A 88:1148-52
Hostomska, Z; Matthews, D A; Davies 2nd, J F et al. (1991) Proteolytic release and crystallization of the RNase H domain of human immunodeficiency virus type 1 reverse transcriptase. J Biol Chem 266:14697-702
Gottlinger, H G; Dorfman, T; Sodroski, J G et al. (1991) Effect of mutations affecting the p6 gag protein on human immunodeficiency virus particle release. Proc Natl Acad Sci U S A 88:3195-9
Davies 2nd, J F; Hostomska, Z; Hostomsky, Z et al. (1991) Crystal structure of the ribonuclease H domain of HIV-1 reverse transcriptase. Science 252:88-95
Hostomsky, Z; Appelt, K; Ogden, R C (1989) High-level expression of self-processed HIV-1 protease in Escherichia coli using a synthetic gene. Biochem Biophys Res Commun 161:1056-63