Macromolecular structure; X-ray crystallography
Our laboratory studies macromolecular structure with an aim toward in-depth understanding of biological activity. Diffraction analysis is our primary research tool, but we also employ theory and other physical and biochemical methods of analysis. The program emphasizes three broad themes: structural biology of specific systems, methodology development, and biophysical principles of conformation, dynamics and assembly.
Most of the macromolecules that we have under study relate to one or more of a few main biological subjects: cell surface interactions and signal transduction, immune response interactions, cellular responses to stress, genetic replication and transcription, carbohydrate recognition, and oxygen transport. Others have been chosen as subject for methodology development or for the analysis of general structural principles. Specific crystalline molecules that we are presently studying include the CD4 T-cell co-receptors, T-cell receptors, MHC molecules, superantigens, stem cell factor, fibroblast growth factor, insulin receptor, lymphocyte kinase, HIV envelope glycoprotein, FHIT, myelin Po, N- cadherin, ribonuclease H, carbamyl phosphate synthetase, UmuD, DnaK, streptavidin, and hemocyanin.
The emphasis in methodology development is on crystallographic phase determination, structure refinement, computational methods, and synchrotron radiation research. Our research in phase determination centers on anomalous scattering -- particularly, on methods for exploiting multiwavelength measurements of anomalous diffraction. Our effort to enhance procedures for stereochemically restrained refinement focuses on an improved treatment of the dynamic characteristics of molecules. A Crystallographic Workbench is being developed for integrated computing. And, with Howard Hughes support, we are developing synchrotron beamlines for macromolecular diffraction studies.
Throughout our work we seek to gain insight into general principles as well as an understanding of specific processes. This is facilitated by methods that allow us to gain structural information in the greatest possible detail and accuracy. General properties that we are addressing include protein dynamics, conformational heterogeneity, metal binding in proteins, determinants of binding strength and specificity, assembly of protein interfaces, molecular symmetry, and bound water structure.
- Peat, T.S., Frank, E.G., McDonald, J.P., Levine, A.S., Woodgate, R. and Hendrickson, W.A. (1996) Structure of the UmuD' protein and its regulation in response to DNA damage. Nature 380, 727-730.
- Fremont, D.H., Hendrickson, W.A., Marrack, P. and Kappler, J. (1996) Structures of an MHC class II molecular with covalently bound single peptides. Science 272, 1001-1004.
- Lima, C.D., Klein, M.G., Weinstein, I.B. and Hendrickson, W.A. (1996) Three-dimensional structure of human protein kinase C interacting protein 1, a member of the HIT family of proteins. Proc. Natl. Acad. Sci USA 93, 5357-5362.
- Zhu, X., Zhao, X., Burkholder, W.F., Gragerov, A., Ogata, C., Gottesman, M.E. and Hendrickson W.A. (1996) Structural analysis of substrate binding by the molecular chaperone DnaK. Science 272, 1605-1614.
- Shapiro, L., Doyle, J.P., Hensley, P., Colman, D.R. and Hendrickson, W.A. (1996) Crystal structure of the extracellular domain from Po, the major structural protein of peripheral nerve myelin. Neuron 17, 435-449.
- Yamaguchi, H. and Hendrickson W.A. (1996) Structural basis for activation of the lymphocyte kinase Lck upon tyrosine phosphorylation. Nature 384, 484-489.