Brian Hales

Brian Hales

brian

Brian  Hales 

Emeritus Professor  - -Biophysical -- Les & Dot Broussard Alumni Association Departmental Professor

Bachelor's Degree(s): Carnegie Institute of Technology, 1966

PhD: University of Minnesota, 1970 Post Doc: Royal Institution of Great Britian, 1970-71

PostDoc: Northwestern University, 1971-73

Phone: (225) 578-4694

Fax: (225) 578-3458

E-mail: bhales@lsu.edu

Office: 327 Choppin Hall

 

Area of Interest

Trace metals have been known for a long time to play many important roles in biology. Our research investigates one of these roles, that being the function of redox active metals and metal clusters in proteins. Metalloproteins constitute some of the most important enzymes known to man. Many of these enzymes are involved with the fundamental reactions needed for life on this planet, and the metal clusters are often the active site. Currently we are investi­gating the mechanism of one of the most important enzymes in Nature, nitrogenase, which catalyzes the fixation or reduction of N2 into ammonia:

 

N2 + 6H+ + 6e- i 2NH3

 

We have used a combination of techniques to probe this mechanism, including those involved with molecular biology, enzymology and spectroscopy. Two recent examples best illustrate the wealth of information that can be obtained.

 

Carbon monoxide is a potent inhibitor of the nitrogenase reaction. In the presence of CO two different EPR signals are generate, one at low pressure (lo-CO, PCO = 0.05 atm) and the second at high pressure (hi-CO, PCO = 0.5 atm). These signals are only generated during enzymatic turnover and thus represent inter­mediate states of catalysis. Using both EPR and ENDOR (electron nuclear double resonance) spectroscopic techniques along with 13C labeled CO and 57Fe label enzyme, we were able to demonstrate that the lo-CO and hi-CO EPR signals arose from CO binding (1 and 2 CO’s, respectively) to FeMo cofactor center of the enzyme. We further revealed that the single CO in lo-CO bound to the cofactor in a bridging fashion while both CO’s in hi-CO exhibited terminal binding.

 

In a related recent study we were able to detect an acetylene adduct binding to the active site during acetylene reduction by nitrogenase. Again, using isotopic labeling (13C and 2H) we demonstrated that this intermediate was a reduced acetylene molecule (C2H2) bridging Fe atoms in the waist region of the FeMo cofactor (see figure) prior to subsequent release and protonation to form ethylene.

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Two views of acetylene bridging across two Fe atoms in the FeMo-cofactor.

 

These results are extremely significant because the represent the first detection of intermediate states of nitrogenase catalysis. We recently expanded our spectroscopic capabilities and are now employing the techniques of MCD (magnetic circular dichroism) spectroscopy and XAS (x-ray absorption spectroscopy, utilizing the LSU CAMD facility) to more completely elucidate the structure of the different intermediate states of nitrogenase as well as other enzymatic reactions.

Awards & Honors

Chair, 2002 Gordon Conference on Nitrogen Fixation, 2002

NIH Biophysical A Study Section, 2001

NIH Special Study Section Reviewer, 1999

Grant Review Panel Member, 1998

Working Group on Hazardous Waste Disposal, 1998

Selected Publications

Zofia Maskos and Brian J. Hales.  Photo Lability of CO Bound to the Cofactor of Mo-nitrogenase from Azotobacker vinelandii.  Journal of Inorganic Biochemistry, 2003, 93, 11-17 

  1. Sorlie, J. Christiansen, B.J. Lemon, J. Peters, D.R. Dean and B. Hales.  Mechanistic Features and Structure of the Nitrogenase MoFe a-Gln195 Protein.  Biochemistry, 2001, 40, 1540-1459
  2. Lee, M. Sorlie, J. Christiansen, R. Song, D.R. Dean, B.J. Hales and B. Hoffman.  Characterization of an Intermediate in the Reduction of Acetylene by the Nitrogenase a-Gln195 MoFe Protein by Q-band EPR and 13C and 1H ENDOR.  J. Amer. Chem. Soc., 2000, 122, 5582-5587
  3. Sorlie, J. Christiansen, D.R. Dean and B.J. Hales.  Detection of a New Radical and FeMo-Cofactor EPR Signal During Acetylene Reduction by the a-H195Q Mutant of Nitrogenase.  J. Amer. Chem. Sco., 1999, 121, 9457-9458

Former Ph.D. Students

Anupam Gupta PhD,Bell Labs
Karen Howard PhD,Procter & Gamble
Melinda Oliver PhD,
Ronald Tittsworth PhD,LSU CAMD
Carol Blanchard PhD,Postdoc
Linda Cameron PhD,Postdoc