E0031

Structural Genomics of Pathogenic Protozoa (SGPP). W. Hol, P. Myler, D. Baker, S. Fields, E. Phizicky, E. Grayhack, M. Dumont, M. Sullivan, C. Mehlin, W. Van Voorhis, F. Buckner, M. Gelb, E. Fan, C. Verlinde, G. DeTitta, J. Luft, D. Meldrum, E. Merritt, M. Berg, T. Earnest, K. Hodgson, L. Brinen, A. Deacon, F. Zucker, Howard Hughes Medical Institute, Depts. of Biochemistry, Chemistry & Genome Sciences, & Medicine, Univ. of Washington, and Seattle Biomedical Research Inst., Seattle, WA; Hauptman-Woodward Medical Inst., Buffalo, NY; Univ. of Rochester, NY; ALS, UC Berkeley, CA; SSRL, Stanford CA.

SGPP focuses on the structures of proteins from Trypanosoma brucei, T. cruzi, Leishmania major and related species, and Plasmodium falciparum, the causative agents of, respectively, sleeping sickness, Chagas’ disease, various forms of leishmaniasis, and malaria. The number of annual deaths due to Trypanosomatid protozoa is in the hundreds of thousands, the morbidity in the tens of millions. P. falciparum is responsible for 300 to 500 million cases of malaria and the deaths of approximately two million young persons per year. The goal of SGPP is to determine the structures of a large number of water-soluble proteins, membrane proteins and protein-protein complexes from these organisms.

Technologies we are developing to high throughput include:

- computational domain prediction

- high-throughput two-hybrid technology to discover protein partners

- several ligation-independent cloning vectors

- vector variants for co-expression of protein pairs

- rapid evaluation of protein expression levels

- rapid generation of single-chain antibodies for co-crystallization

- synthesis of tripartite, low molecular weight co-crystallants

- high-throughput crystallization and crystal-imaging robotics

- improved beam line technologies for rapid data collection

- bioinformatics and informatics capability for data storage and mining

Work supported in part by NIH Grant 1-P50- GM64655-01