The long-term goal of this work is to understand the mechanism of coupling in F1F0 ATP synthase. Most organisms synthesize ATP via F1F0 ATP synthases so these enzymes are largely responsible for the central function of energy metabolism. In humans, failures in energy metabolism are associated with aging and a variety of inherited disorders. For example, the disease in certain individuals with Leigh's Syndrome is directly attributed to mutations affecting F1F0 ATP synthase. Prokaryotic and eucaryotic F1F0 ATP synthases share common structures and mechanisms. Therefore, the Escherichia coli F1F0 ATP synthase has been chosen as a model. The E. coli system offers a well-characterized enzyme and the use of the broadest range of molecular genetic techniques. Recently, much attention has been focussed on the two stalk structures linking the F1 and F0 sectors of the enzyme. The central stalk is thought to rotate within the enzyme complex during catalysis, while the peripheral stalk serves as a stator holding the bulk of F1 in place. This has led to the widely held view that the peripheral stalk is a rigid static structural feature, but this interpretation is not consistent with all experimental observations. It is hypothesized that the peripheral stalk plays a dynamic role in coupling. The following Specific Aims are proposed to investigate this concept: 1) determining the length of the peripheral stalk in enzymes with deletions in the b subunits; 2) studying the site of interaction between the a and b subunits; 3) investigating the dynamics of the b subunit interaction with F1 by looking for evidence of transient dissociation of the peripheral stalk; 4) studying the preferred intermediates in the assembly of F1F0 ATP synthase.

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National Institute of General Medical Sciences (NIGMS)
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Special Emphasis Panel (ZRG1-SSS-A (03))
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Preusch, Peter C
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University of Florida
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Bhatt, Deepa; Cole, Stephanie P; Grabar, Tammy Bohannon et al. (2005) Manipulating the length of the b subunit F1 binding domain in F1F0 ATP synthase from Escherichia coli. J Bioenerg Biomembr 37:67-74
Grabar, Tammy Bohannon; Cain, Brian D (2004) Genetic complementation between mutant b subunits in F1F0 ATP synthase. J Biol Chem 279:31205-11
Grabar, Tammy Bohannon; Cain, Brian D (2003) Integration of b subunits of unequal lengths into F1F0-ATP synthase. J Biol Chem 278:34751-6
Hardy, Andrew W; Grabar, Tammy Bohannon; Bhatt, Deepa et al. (2003) Mutagenesis studies of the F1F0 ATP synthase b subunit membrane domain. J Bioenerg Biomembr 35:389-97
Yi, Liang; Jiang, Fenglei; Chen, Minyong et al. (2003) YidC is strictly required for membrane insertion of subunits a and c of the F(1)F(0)ATP synthase and SecE of the SecYEG translocase. Biochemistry 42:10537-44
Gardner, J L; Cain, B D (1999) Amino acid substitutions in the a subunit affect the epsilon subunit of F1F0 ATP synthase from Escherichia coli. Arch Biochem Biophys 361:302-8
Hartzog, P E; Cain, B D (1994) Second-site suppressor mutations at glycine 218 and histidine 245 in the alpha subunit of F1F0 ATP synthase in Escherichia coli. J Biol Chem 269:32313-7
Stack, A E; Cain, B D (1994) Mutations in the delta subunit influence the assembly of F1F0 ATP synthase in Escherichia coli. J Bacteriol 176:540-2
Hartzog, P E; Cain, B D (1993) Mutagenic analysis of the a subunit of the F1F0 ATP synthase in Escherichia coli: Gln-252 through Tyr-263. J Bacteriol 175:1337-43
McCormick, K A; Deckers-Hebestreit, G; Altendorf, K et al. (1993) Characterization of mutations in the b subunit of F1F0 ATP synthase in Escherichia coli. J Biol Chem 268:24683-91

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