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William Furey, PhD

Biocrystallography Laboratory, PO Box 12055
VA Medical Center
Pittsburgh, PA 15240
Phone: 412-607-3106
Fax: 412-688-6945


BA (Chemistry), Rutgers, The State University of New Jersey, 1974
PhD (Physical Chemistry- Crystallography) Rutgers, The State University of New Jersey, 1977
Postdoctoral Fellow, Department of Crystallography, University of Pittsburgh, 1981
Headshot of William Furey, PhD


Research Areas

Dr. Furey’s research involves the structure determination and analysis of large biological molecules and complexes by x-ray crystallography, and correlating the results with known functions. The work currently focuses on thiamin (vitamin B1) dependent enzymes and cell cycle regulating enzymes, as well as crystallographic methods development. Results of these studies could lead to development of therapeutic agents directed against pathogenic organisms, and anti-cancer drugs.


The pyruvate dehydrogenase multienzyme complex (PDHc, MW 4.7 million Daltons, 60 protein subunits & 60 active sites for the E. coli version) is present in most organisms and is critical for carbohydrate metabolism where it converts pyruvate, the product of glycolysis, to acetyl-CoA via a complicated process of substrate channeling within the confines of the complex. Structural analyses of the complex and its three major enzymatic components E1 (24 copies), E2 (24 copies), & E3 (12 copies) are underway, and Dr. Furey has already determined high resolution crystal structures for some of the components and reaction intermediates from the E. coli version.


The E1 components are rate determining and require thiamin diphosphate as a cofactor, but must interact with a flexible segment [lipoyl domain (LD) and associated lipoamide side chain] on an E2 to transfer the first reaction product, an acetyl group, to the E2 active site. The acetyl group is then transferred to co-enzyme A within the E2 active site, and the product acetyl-CoA is released. The E2 bound lipoamide group (now reduced) then moves to an E3 (FAD dependent) active site, where it is oxidized to restore the initial conditions. Binding of the flexible segment to E1 and E3 subunits is mediated by additional binding to a peripheral subunit-binding domain (PSBD), shown bound to E1.



Mechanistic details regarding the catalytic reactions in each active site are sought, as well as identifying structural aspects critical for assembly of the individual components to form the complete multienzyme complex. Specific mutations in some of the components are associated with hereditary diseases in humans, and detailed analyses of the structure-function relationships may suggest development of plausible therapeutic agents to counter the effects of the mutations. Additionally, given the critical nature of this system in overall energy production for cellular function, development of inhibitors binding at any of the catalytic sites, or at sites disrupting protein-protein assembly, may considerably weaken or kill the organism. Lack of appreciable sequence homology between PDHc’s from humans and pathogenic bacteria therefore suggests that effective, pathogen specific antibacterial agents may be developed.


Early expression or over expression of Cdc25 proteins can cause the cell to prematurely progress leading to oncogenic effects, making these enzymes exciting targets for anti-cancer drug development. As part of a collaborative effort with Dr. John Lazo’s group, several potent inhibitors of Cdc25 proteins have been discovered, and structural analysis of their complexes with the enzymes are underway to reveal both where and how these inhibitors function. Dr. Furey has crystallized the catalytic domain of Cdc25b and determined its high-resolution structure. His group is currently co-crystallizing the catalytic domain with several inhibitors, as a step towards development of effective anti-cancer agents via structure-based drug design procedures.


In collaboration with the Hauptman-Woodward Institute for Medical Research, Dr. Furey's group is developing new computational methods for solving macromolecular crystal structures by automated techniques. This work involves creating and developing a software package BnP, which is a merging of the PHASES package developed in the Furey lab, and the SnB package developed in Buffalo. A simple, graphical user interface is developed to enable automatic creation of an interpretable electron density map starting from observed x-ray diffraction data, with only a few mouse clicks and text field entries required. This will invoke automatic scaling of data, determination of heavy atom/anomalous scatterer sites, refinement and validation of sites, calculation of protein phases, phase refinement, and phase improvement via solvent flattening/negative density truncation. A few more mouse clicks enable automated building of a complete or nearly complete model by interfacing with other externally developed software. The idea is to make it simple for novices to determine good quality crystal structures, while enhancing the productivity of more sophisticated users as well.

Journal Articles

Koharudin L, W Furey and A Gronenborn.  Novel fold and carbohydrate specificity of the potent anti-HIV cyanobacterial lectin from oscillatoria agardhii.  J Biol Chem 286:1588-1597, 2011.
Nemeria N, P Arjunan, K Chandrasekhar, M Mossad, K Tittmann, W Furey and F Jordan.  Communication between thiamin cofactors in the Escherichia coli pyruvate dehydrogenase complex E1 component active center:  Evidence for a "direct pathway" between the 4'-aminopyridine N1' atoms.  J Biol Chem 285:11197-11209, 2010.
Matei E, A Zheng, W Furey, J Rose, C Aiken and A Gronenborn.  Anti-HIV activity of defective cyanovirin-N mutants is restored by dimerization.  J Biol Chem 285:13057-13065, 2010.
Jordan F, P Arjunan, S Kale, N Nemeria and W Furey.  Multiple roles of mobile active center loops in the E1 component of the Escherichia coli pyruvate dehydrogenase complex:  Linkage of protein dynamics to catalysis.  J Mol Cat B:  Enzymatic 61:14-22, 2009.
Kale S, G Ulas, J Song, G Brudvig, W Furey and F Jordan. Efficient coupling of catalysis and dynamics in the E1 component of escherichia coli pyruvate dehydrogenase multienzyme complex. Proc Natl Acad Sci 105(4):1158-1163, 2008.
Arjunan P, M Sax, A Brunskill, K Chandrasekhar, N Nemeria, S Zhang, F Jordan and W Furey. A thiamin-bound, pre-decarboxylation reaction intermediate analogue in the pyruvate dehydrogenase E1 subunit induces large-scale disorder-to-order transformations in the enzyme and reveals novel structural features in the covalently bound adduct. J Biol Chem 281:15296-15303, 2006.
Jordan F, N Nemeria, S Zhang, Y Yan, P Arjunan and W Furey. Dual catalytic apparatus of the thiamin diphosphate coenzyme: Acid-base via the 1’.4’ iminopyrimidine tautomer along with its electrophilic role. J Am Chem Soc 125:12732-12738, 2003.
Nemeria N, P Arjunan, A Brunskill, F Sheibani, W Wei, Y Yan, S Zhang, F Jordan and W Furey. Histidine 407, a phantom residue in the E1 subunit of the Escherichia coli pyruvate dehydrogenase complex, activates reductive acetylation of lipoamide on the E2 subunit. An explanation for conservation of active sites between the E1 subunit and transketolase. Biochemistry 41(52):15459-15467, 2002.
Arjunan P, N Nemeria, A Brunskill, K Chandrasekhar, M Sax, Y Yan, F Jordan, J Guest and W Furey. Structure of the pyruvate dehydrogenase multienzyme complex E1 component from Escherichia coli at 1.85Å resolution. Biochemistry 41(16):5213-5221, 2002.
Furey W and S Swaminathan. PHASES-95: A Program Package for Processing and Analysing Diffraction Data From Macromolecules in Methods in Enzymology: Macromolecular Crystallography, Part B, Vol 277, eds. C. Carter & R. Sweet, Academic Press, Orlando, Fl., pp 590-620, 1997.

Sponsored Research

Pyruvate Dehydrogenase Assembly, Structure & Function - 7/1/2017 - 6/30/2021
NIH - R01 GM121469