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Stephen Everse, Ph.D.
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Work Address
Biochemistry, Given B418A
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Stephen.Everse@uvm.edu
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Summary
Background:
Dr. Everse received his Ph.D. in Chemistry from the University of California, San Diego in 1995. His postdoctoral work at UCSD focused on obtaining a structural understanding of the fibrinogen molecule. His work resulted in the X-ray crystallographic structures of the fibrinogen fragment D and the fibrin fragment double-D. He joined the Biochemistry department in the Fall of 1998 as part of the HHMI structural biology initiative at UVM. HHMI provided for a state-of-the-art X-ray facility including a rotating anode generator, two MAR345 detectors, two cryo-cooling systems and some of our computer resources. A DOE EPSCoR award to Dr. Susan Wallace (Microbiology & Molecular Genetics) added incubators, a crystallization robot, a robotic viewing station, and additional computer resources for structural determination.
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Research Overview
Research Interests:
Physical Biochemistry
Structural Biology
Project Description:
His laboratory takes a crystallographic approach to address five fundamental questions: How is protein function defined by structure? What determines and mediates protein-protein and protein-membrane interactions? How does information get passed from one protein to another? How do protein cofactors modulate enzymes? How does structure prescribe the binding affinity of a metal? Three model systems are being studied to explore these basic questions:
Factor V is a large (330 kDa), single chain, coagulation cofactor which is proteolytically activated by other coagulation factors. As a start to providing a real structural understanding of the cofactors we, in collaboration with Dr. Kenneth G. Mann (Biochemistry), have solved the structure of bovine factor Vai (Figure 1). While attempting to crystallize both factor Va and the prothrombinase complex we have been building and refining models of the prothrombinase complex.
Transferrin (80 kDa) is a glycoprotein with two homologous lobes. Each lobe consists of two domains that form a deep cleft which bind a single Fe (III) ion in conjunction with the concomitant binding of a carbonate ion in a pH dependent manner (Figure 2A). In collaboration with Dr. Anne B. Mason (Biochemistry) we have been solving structures of mutants of the N-lobe of human serum transferrin (hTF) in order to understand the iron uptake and release properties of this protein. In addition we continue efforts to determine structures of diferric and monoferric hTFs.
Thioredoxin reductase is part of the thioredoxin system, an antioxidant system responsible for protecting the cell against oxidative stress. In collaboration with Dr. Robert Hondal (Biochemistry) we have been solving structures of thioredoxin reductases (Figure 2B). Our goal is to gain a structural insight into the mechanism and the role of selenocysteine.
Figure 1: Structure of bovine factor Vai (Adams et al. 2004 PNAS 101:8918-23).
Figure 2: Oxalate (orange) in the iron binding cleft of the N-lobe of human serum transferrin
(Halbrooks et al. 2004 JMB 339:217-26).
Alignment of the tetrapeptide SCCS(ox) in the structure of TR from Drosophila. The tetrapeptide structures (B: C+ conformation and C: T- conformation determined by NMR spectroscopy) were modeled in the active site of TR providing a structural basis for our mechanism (for details see Eckenroth et al 2007 Biochemistry 46:4694-705).
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Publications
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- Everse SJ, Doublie S. (2006). Abstract Crystallographic software: a sustainable resource for the community. Methods Mol Biol., 364:273-8.
- Adams TE, Hockin MF, Mann KG, and Everse SJ. (2004). The crystal structure of activated protein C-inactivated bovine factor Va: Implications for cofactor function. Proc Natl Acad Sci USA,101(24):8918-23. Epub 2004 Jun 07 View the structure at PDB
- Halbrooks PJ, Mason AB, Adams TE, Briggs SK, Everse SJ. (2004). The oxalate effect on release of iron from human serum transferrin explained. J Mol Biol., 339(1):217-26. View the structure at PDB
- Adams TE, Everse SJ, Mann KG. (2003). Predicting the pharmacology of thrombin inhibitors. J Thromb Haemost.,1(5):1024-7.
- Halbrooks PJ, He QY, Briggs SK, Everse SJ, Smith VC, MacGillivray RT, and Mason AB. (2003). Investigation of the Mechanism of Iron Release from the C-Lobe of Human Serum Transferrin: Mutational Analysis of the Role of a pH Sensitive Triad. Biochemistry, 42(13):3701-7
- Adams TE, Mason AB, He QY, Halbrooks PJ, Briggs SK, Smith VC, MacGillivray RT, and Everse SJ. (2003). The Position of Arginine 124 Controls the Rate of Iron Release from the N-lobe of Human Serum Transferrin. A STRUCTURAL STUDY. J Biol Chem., 278(8):6027-33. View the structure at PDB
- Yang Z, Spraggon G, Pandi L, Everse SJ, Riley M, Doolittle RF. (2002). Crystal structure of fragment D from lamprey fibrinogen complexed with the peptide Gly-His-Arg-Pro-amide. Biochemistry, 41(32):10218-24. View the structure at PDB
- Hockin MF, Jones KC, Everse SJ, Mann KG. (2002). A model for the stoichiometric regulation of blood coagulation. J Biol Chem., 277(21):18322-33.
- Everse SJ. (2002). New insights into fibrin (ogen) structure and function. Vox Sang., 83 Suppl 1:375-82.
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All Everse Publications
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