Macromolecular crystal growth; X-ray diffraction analysis of protein, nucleic acid and virus crystals; Atomic Force Microscopy studies of biological structures, immunoglobulin structure, virus structure and assembly
2003-2004 Recipient of Fankuchen Award at 2004 ACA Meeting, Chicago, IL
1999 Recipient of NASA Medal for Exceptional Scientific Achievement
Research is divided among a number of areas focused ultimately on the determination and description of biological structure, principally at the molecular level, but extending to the organelle and cellular as well. The primary techniques employed are X-ray crystallography and Atomic Force Microscopy (AFM). In support of the application of these biophysical methods we employ a wide range of conventional biochemical, as well as molecular biological techniques. Of chief interest in terms of molecular structure are intact immunoglobulins, whose dynamics we hope to reveal through a series of static, X-ray derived images, and small spherical viruses for which we are attempting to define the molecular structures and mechanisms involved in assembly. To promote our research in structure determination, a major interest is in the growth of macromolecular crystals. Using AFM and QELS we are attempting to elucidate the phase transition phenomenon leading to nucleation, and the mechanisms, kinetic factors, and thermodynamic parameters fundamental to the crystal growth process. Among the diverse approaches utilized, is crystallization of macromolecules in microgravity, including missions aboard the U.S. Space Shuttle and the International Space Station. These experiments allow us, in many instances, to improve the quality of crystals used in our analyses and hence, the precision of the molecular structures derived from those crystals. Most recently we have undertaken investigations of living cells, animal viruses, bacteriophage, giant viruses such as PBCV-1 and mimivirus, and virus infected cells using AFM. These have included herpes simplex virus, vaccinia, and several retroviruses including mouse leukemic virus and human immunodeficiency virus.
Investigation of bacteriophage T4 by atomic force microscopy.
Kuznetsov YG, Chang SC, McPherson A.
Bacteriophage. 2011 May;1(3):165-173. Epub 2011 May 1.
Kuznetsov, Y.G., Xiao, C., Sun, S., Raoult, D., Rossmann, M. and McPherson, A. Atomic Force Microscopy investigation of the giant mimivirus. Virology 404, 127-37 (2010).
Nitta, T., Kuznetsov, Y.G., McPherson, A. and Fan, H. Murine Leukemic Virus Glycosylated Gag (gPr80gag) Facilitates Interferon-Sensitive Virus Release through lipid rafts. Proc. Nat. Acad. Sci. USA 107, 1190-1195 (2010).
Larson, S.B., Day, J. and McPherson, A. X-ray crystallographic analysis of pig pancreatic alpha-amylase with limit dextrin, oligosaccharide and alpha – cyclodextrin. Biochemistry 49, 3101-3115 (2010).
Xiao, C., Kuznetsov, Y.G., Sun, S., Hafenstein, S.L., Kostyuchenko, V.A., Chipman, P.R., Suzan-Monti, M., Raoult, D., McPherson, A. and Rossmann, M.G. Structural studies of the giant Mimivirus. PLoS Biology 7 (4) 958-966 (2009).
McPherson, A. Introduction to protein crystallization. In Membrane Protein Crystallization: Current Topics in Membranes, vol. 63. Ed. L. DeLucas. Elsevier, Co. 5-3 (2009).
McPherson, A. Introduction to Macromolecular Crystallography. 2nd edition, John Wiley Co., NY (2009).
Larson, S.B., Day, J.S., Nguyen, C., Cudney, R. and McPherson, A. Pig heart citrate synthase refined at 1.7 Å resolution. Acta Cryst. F65, 430-434 (2009).
Larson, S.L., Day, J.S., Nguyen, C., Cudney, R. and McPherson, A. High resolution structure of a complex between proteinase K and digalacturonic acid. Acta Cryst. F65, 192-198 (2008).
Kuznetsov, Yu. G., Gershon, P. D. and McPherson, A. Atomic Force Microscopy Investigation of Vaccinia Virus Structure. J. Virology 82 (15) 7551-7566 (2008).
Larson, S.B., Day, J.S., Nguyen, C., Cudney, B. and McPherson, A. Progress in the Development of an Alternative Approach to Macromolecular Crystallization. Crystal Growth and Design 8 (8) 3038-3052 (2008).
Kuznetsov, Yu. G., Daijogo, S., Zhou, J., Semler, B.L. and McPherson, A. Atomic Force Microscopy Analysis of Icosahedral Virus RNA. J. Mol. Biol. 347, 41-52 (2005).
Makino, D., Henschen-Edman, A. H., Larson, S. B., Day, J. S., McPherson, A. Bence Jones KWR protein structures determined by X-ray crystallography. Acta Cryst. 63 780-92 (2007).
Kuznetsov, Yu G., Ulbrich, P., Haubova, S., Ruml, T. and McPherson, A. Atomic Force Microscopy Investigation of Mason - Pfizer Monkey Virus and Human Immunodeficiency Virus Type 1 Reassembled Particles. Virology 350(2) 434-446 (2007).
McPherson, A. and Cudney, B. Searching for Silver Bullets: An Alternative Strategy for Crystallizing Macromolecules. J. Struct. Biol., 156, 387-406 (2006).
McPherson, A. and Kuznetsov, Yu. G. Atomic Force Microscopy Investigation of Turnip Yellow Mosaic Virus Capsid Disruption and RNA Extrusion. Virology 352, 329-337 (2006).
Makino, D., Day, J. S., Larson, S. B., McPherson, A. Investigation of RNA Structure in Satellite Panicum Mosaic Virus. Virology 351, 420-431 (2006).
Freddolino, P. L., Arkhipov, A.S., Larson, S. B., McPherson, A. and Schulten, K. Molecular Dynamics Simulations of the Complete Satellite Tobacco Mosaic Virus. Structure 14, 1-13 (2006).
294. Larson, S. B., Day, J. S., Glasser, S., Braslawsky, G. and McPherson, A. The structure of an antitumor CH2 – domain deleted humanized antibody. J. Mol. Biol. 348, 1177–1190 (2005).
Larson, S. B., Lucas, R. W., Greenwood, A. and McPherson, A. The RNA of turnip yellow mosaic virus exhibits icosahedral order. Virology 334, 245-254 (2005).
Structural Biology and Molecular Biophysics