The specific aims of the proposed research are: 1) to finalize the structure of the functionalized non-heme iron center (H cluster) that is uniquely involved in both substrate (H2) activation and electron transfer; 2) to analyze the function of the H cluster and two ferredoxin- type clusters by specific probes utilizing biophysical techniques; 3) to attempt to isolate the gene product of the putative hydrogenase gene, hyd lambda from Desulfovibrio vulgaris which is suspected to be a new-type of (Fe) hydrogenase in Desulfovibrio and may resolve the problem of the number and types of (Fe) hydrogenases; 4) to establish a DNA transfer system with D. vulgaris using a wide host range plasmid and generate (Fe) hydrogenase negative mutants in order to obtain biosynthesis of active (Fe) hydrogenase and 5) to investigate the role of the small subunit and its putative signal peptide in the periplasmic localization of the (Fe) hydrogenase with the goal of determining how the large subunit that lacks a signal peptide is translocated across the cytoplasmic membrane.

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Project (R01)
Project #
5R01GM034903-07
Application #
3286741
Study Section
Microbial Physiology and Genetics Subcommittee 2 (MBC)
Project Start
1985-09-05
Project End
1995-07-31
Budget Start
1992-08-01
Budget End
1993-07-31
Support Year
7
Fiscal Year
1992
Total Cost
Indirect Cost
Name
University of Georgia
Department
Type
Schools of Arts and Sciences
DUNS #
City
Athens
State
GA
Country
United States
Zip Code
30602
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DerVartanian, M E; Menon, N K; Przybyla, A E et al. (1996) Electron paramagnetic resonance (EPR) studies on hydrogenase-1 (HYD1) purified from a mutant strain (AP6) of Escherichia coli enhanced in HYD1. Biochem Biophys Res Commun 227:211-5
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DerVartanian, D V (1994) Desulforubidin: dissimilatory, high-spin sulfite reductase of Desulfomicrobium species. Methods Enzymol 243:270-6
Le Gall, J; Payne, W J; Chen, L et al. (1994) Localization and specificity of cytochromes and other electron transfer proteins from sulfate-reducing bacteria. Biochimie 76:655-65
Patil, D S (1994) Nickel-iron-selenium hydrogenase. Methods Enzymol 243:68-94
Menon, N K; Robbins, J; Der Vartanian, M et al. (1993) Carboxy-terminal processing of the large subunit of [NiFe] hydrogenases. FEBS Lett 331:91-5
Picarra-Pereira, M A; Turner, D L; LeGall, J et al. (1993) Structural studies on Desulfovibrio gigas cytochrome c3 by two-dimensional 1H-nuclear-magnetic-resonance spectroscopy. Biochem J 294 ( Pt 3):909-15
Przybyla, A E; Robbins, J; Menon, N et al. (1992) Structure-function relationships among the nickel-containing hydrogenases. FEMS Microbiol Rev 8:109-35

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