Abstract

This work involves the general characterization of proteins involved in bacterial respiration. The specific subjects of this proposal are the cytochrome claa3 and Rieske iron/sulfur protein isolated from the extremely thermophilic aerobe, Thermus thermophilus. The overall objective of the work is to provide physical insight on the mechanism of coupled proton translocation and electron transfer. The methods used include general techniques of protein purification and characterization; those of recombinant DNA technology to isolate and sequence the genes of interest; and physical methods including EPR, ENDOR, Mossbauer, and resonance Raman spectroscopies. Kinetic studies are designed for the detection and characterization of intermediates in dioxygen reduction, and for the detection of spectral perturbations of vesicular cytochrome aa3 under the influence of a polarizing electric field. Guided by the belief that understanding mechanisms in respiration will require detailed three-dimensional structures, we are making a serious effort to crystallize these proteins.

  Funding Agency

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Project (R01)
Project #
5R01GM035342-06
Application #
3287935
Study Section
Physical Biochemistry Study Section (PB)
Project Start
1984-12-01
Project End
1992-11-30
Budget Start
1989-12-01
Budget End
1990-11-30
Support Year
6
Fiscal Year
1990
Total Cost
Indirect Cost

  Institution

Name
Los Alamos National Lab
Department
Type
Organized Research Units
DUNS #
City
Los Alamos
State
NM
Country
United States
Zip Code
87545

  Publications

McDonald, William; Funatogawa, Chie; Li, Yang et al. (2014) Conserved glycine 232 in the ligand channel of ba3 cytochrome oxidase from Thermus thermophilus. Biochemistry 53:4467-75
McDonald, William; Funatogawa, Chie; Li, Yang et al. (2013) Ligand access to the active site in Thermus thermophilus ba(3) and bovine heart aa(3) cytochrome oxidases. Biochemistry 52:640-52
Luna, V Mitch; Fee, James A; Deniz, Ashok A et al. (2012) Mobility of Xe atoms within the oxygen diffusion channel of cytochrome ba(3) oxidase. Biochemistry 51:4669-76
Neehaul, Yashvin; Chen, Ying; Werner, Carolin et al. (2012) Electrochemical and infrared spectroscopic analysis of the interaction of the Cu(A) domain and cytochrome c(552) from Thermus thermophilus. Biochim Biophys Acta 1817:1950-4
Chang, Hsin-Yang; Choi, Sylvia K; Vakkasoglu, Ahmet Selim et al. (2012) Exploring the proton pump and exit pathway for pumped protons in cytochrome ba3 from Thermus thermophilus. Proc Natl Acad Sci U S A 109:5259-64
Egawa, Tsuyoshi; Chen, Ying; Fee, James A et al. (2012) The rate-limiting step in O(2) reduction by cytochrome ba(3) from Thermus thermophilus. Biochim Biophys Acta 1817:666-71
Liu, Bin; Zhang, Yang; Sage, J Timothy et al. (2012) Structural changes that occur upon photolysis of the Fe(II)(a3)-CO complex in the cytochrome ba(3)-oxidase of Thermus thermophilus: a combined X-ray crystallographic and infrared spectral study demonstrates CO binding to Cu(B). Biochim Biophys Acta 1817:658-65
Tiefenbrunn, Theresa; Liu, Wei; Chen, Ying et al. (2011) High resolution structure of the ba3 cytochrome c oxidase from Thermus thermophilus in a lipidic environment. PLoS One 6:e22348
Smirnova, Irina; Reimann, Joachim; von Ballmoos, Christoph et al. (2010) Functional role of Thr-312 and Thr-315 in the proton-transfer pathway in ba3 Cytochrome c oxidase from Thermus thermophilus. Biochemistry 49:7033-9
Moënne-Loccoz, Pierre; Fee, James A (2010) Biochemistry. Catalyzing NO to N2O in the nitrogen cycle. Science 330:1632-3

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