Inhibition of ATPase activity of E. coli F1 by the water-soluble carbodiimide EDC is due to modification of several carboxyls in the .beta. subunit

Lotscher, Hans Ruedi; DeJong, Catherina; Capaldi, Roderick A.

doi:10.1021/bi00313a019

Cited by 52 publications

(34 citation statements)

References 36 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…The binding site for NCCD is believed to be the same as that for DCCD, i.e., Asp61 of subunit c, as NCCD's binding to the ATP synthase is prevented by DCCD (24). Moreover, the mutant of (236). Incorporation of about 13 mol of EDC/mol F 1 (E. coli) leads to 95% inhibition of ATPase activity.…”

Section: Carboxyl Group Modifiersmentioning

confidence: 99%

“…Incorporation of about 13 mol of EDC/mol F 1 (E. coli) leads to 95% inhibition of ATPase activity. Here, two-thirds of the bound EDC is bound to ␤ subunits, where it modifies multiple sites in a short segment (residues 162 to 194) (E. coli sequence) (236). EDC also promotes formation of intersubunit crosslinks between subunits ␤ and ε.…”

Section: Carboxyl Group Modifiersmentioning

confidence: 99%

See 1 more Smart Citation

ATP Synthase and the Actions of Inhibitors Utilized To Study Its Roles in Human Health, Disease, and Other Scientific Areas

Hong

Pedersen

2008

Microbiol Mol Biol Rev

275

302

View full text Add to dashboard Cite

SUMMARY ATP synthase, a double-motor enzyme, plays various roles in the cell, participating not only in ATP synthesis but in ATP hydrolysis-dependent processes and in the regulation of a proton gradient across some membrane-dependent systems. Recent studies of ATP synthase as a potential molecular target for the treatment of some human diseases have displayed promising results, and this enzyme is now emerging as an attractive molecular target for the development of new therapies for a variety of diseases. Significantly, ATP synthase, because of its complex structure, is inhibited by a number of different inhibitors and provides diverse possibilities in the development of new ATP synthase-directed agents. In this review, we classify over 250 natural and synthetic inhibitors of ATP synthase reported to date and present their inhibitory sites and their known or proposed modes of action. The rich source of ATP synthase inhibitors and their known or purported sites of action presented in this review should provide valuable insights into their applications as potential scaffolds for new therapeutics for human and animal diseases as well as for the discovery of new pesticides and herbicides to help protect the world's food supply. Finally, as ATP synthase is now known to consist of two unique nanomotors involved in making ATP from ADP and Pi, the information provided in this review may greatly assist those investigators entering the emerging field of nanotechnology.

show abstract

Section: Carboxyl Group Modifiersmentioning

confidence: 99%

Section: Carboxyl Group Modifiersmentioning

confidence: 99%

ATP Synthase and the Actions of Inhibitors Utilized To Study Its Roles in Human Health, Disease, and Other Scientific Areas

Hong

Pedersen

2008

Microbiol Mol Biol Rev

275

302

View full text Add to dashboard Cite

show abstract

“…The identity of the subunits involved in cross-links was revealed by use of monoclonal antibodies on Western blots (Mendel-Hartvig and Capaldi, 1991). ATPase activity was measured in a regenerating system according to Lötscher et al (1984). Protein concentrations were determined with the BCA protein assay from Pierce.…”

Section: Construction Of Plasmids Containingmentioning

confidence: 99%

Nucleotide-dependent Movement of the ε Subunit between α and β Subunits in the Escherichia coli F1F0-type ATPase

Aggeler

Capaldi

1996

Journal of Biological Chemistry

Self Cite

View full text Add to dashboard Cite

Mutants of ECF 1 -ATPase were generated, containing cysteine residues in one or more of the following positions: ␣Ser-411, ␤Glu-381, and ⑀Ser-108, after which disulfide bridges could be created by CuCl 2 induced oxidation in high yield between ␣ and ⑀, ␤ and ⑀, ␣ and ␥, ␤ and ␥ (endogenous Cys-87), and ␣ and ␤. All of these cross-links lead to inhibition of ATP hydrolysis activity. In the two double mutants, containing a cysteine in ⑀Ser-108 along with either the DELSEED region of ␤ (Glu-381) or the homologous region in ␣ (Ser-411), there was a clear nucleotide dependence of the cross-link formation with the ⑀ subunit. In ␤E381C/⑀S108C the ␤-⑀ cross-link was obtained preferentially when Mg 2؉ and ADP ؉ P i (addition of MgCl 2 ؉ ATP) was present, while the ␣-⑀ cross-link product was strongly favored in the ␣S411C/⑀S108C mutant in the Mg 2؉ ATP state (addition of MgCl 2 ؉ 5-adenylyl-␤,␥-imidodiphosphate). In the triple mutant ␣S411C/␤E381C/⑀S108C, the ⑀ subunit bound to the ␤ subunit in Mg 2؉ -ADP and to the ␣ subunit in Mg There has been significant recent progress in determining the structure of the F 1 part of the F 1 F 0 -type ATP synthase, a key enzyme in oxidative phosphorylation and photo-phosphorylation (Senior 1988(Senior , 1990Boyer, 1993). The F 1 , which can be detached from the F 0 and studied separately, is a complex of five different types of subunits ␣, ␤, ␥, ␦, and ⑀ in the molar ratio 3:3:1:1:1. Electron microscopy has shown that the ␣ and ␤ subunits are hexagonally arranged, and alternate around a central cavity in which the ␥ subunit is located (Gogol et al. 1989a(Gogol et al. , 1989bBoekema and Böttcher, 1992). Biochemical studies place the ␦ and ⑀ subunits (nomenclature for the Escherichia coli enzyme) at the bottom of the ␣ 3 ␤ 3 ␥ core complex (Beckers et al., 1992;Dallmann et al., 1992;Capaldi et al., 1994Capaldi et al., , 1995 in the stalk region which is a 40 -45-Å long structure that links the F 1 to the F 0 part (Gogol et al., 1987;Lü cken et al., 1990). The recently published high resolution structure of a major part of the F 1 molecule confirms the above described arrangement of the ␣, ␤, and ␥ subunits relative to one another and adds important details (Abrahams et al., 1994). The ␣ and ␤ subunits have a similar overall fold, each made up of three domains: an NH 2 -terminal, predominantly ␤-sheet domain, a nucleotide-binding domain of 9 ␤ strands and 9 ␣ helices, and a COOH-terminal, predominantly ␣-helical domain, that provides the binding site for the ⑀ subunit, probably the ␦ subunit, as well as the b subunit of the F 0 . The part of the ␥ subunit within the ring of ␣ and ␤ subunits is arranged as two ␣ helices: a long COOH-terminal helix extending from the top NH 2 -terminal domain of the ␣ and ␤ subunits into the stalk region, and a short NH 2 -terminal helix running from the catalytic site domain into the stalk. These two ␣ helices form a coiled coil. A third short ␣-helix of the ␥ subunit (residues 82-99 in the E. coli enzyme) is inclined at about 45°to the two larger helices...

show abstract

“…The ⑀ subunit is thought to inhibit by reducing the rate of product release (10), the rate-limiting step in hydrolysis. It is known to bind to isolated ␥ subunit (11), but can be cross-linked to ␣, ␤, and ␥ subunits (12)(13)(14)(15).…”

mentioning

confidence: 99%

Alanine-scanning Mutagenesis of the ∊ Subunit of the F1-F0 ATP Synthase from Escherichia coli Reveals Two Classes of Mutants

Xiong

Vik

1995

Journal of Biological Chemistry

View full text Add to dashboard Cite

Alanine-scanning mutagenesis was applied to the ⑀ subunit of the F 1 -F 0 ATP synthase from E. coli. Nineteen amino acid residues were changed to alanine, either singly or in pairs, between residues 10 and 93. All mutants, when expressed in the ⑀ deletion strain XH1, were able to grow on succinate minimal medium. Membranes were prepared from all mutants and assayed for ATPdriven proton translocation, ATP hydrolysis ؎ lauryldiethylamine oxide, and sensitivity of ATPase activity to N,N-dicyclohexylcarbodiimide (DCCD). Most of the mutants fell into 2 distinct classes. The first group had inhibited ATPase activity, with near normal levels of membrane-bound F 1 , but decreased sensitivity to DCCD. The second group had stimulated ATPase activity, with a reduced level of membrane-bound F 1 , but normal sensitivity to DCCD. Membranes from all mutants were further characterized by immunoblotting using 2 monoclonal antibodies. A model for the secondary structure of ⑀ and its role in the function of the ATP synthase has been developed. Some residues are important for the binding of ⑀ to F 1 and therefore for inhibition. Other residues, from Glu-59 through Glu-70, are important for the release of inhibition by ⑀ that is part of the normal enzyme cycle.

show abstract

Inhibition of ATPase activity of E. coli F1 by the water-soluble carbodiimide EDC is due to modification of several carboxyls in the .beta. subunit

Cited by 52 publications

References 36 publications

ATP Synthase and the Actions of Inhibitors Utilized To Study Its Roles in Human Health, Disease, and Other Scientific Areas

ATP Synthase and the Actions of Inhibitors Utilized To Study Its Roles in Human Health, Disease, and Other Scientific Areas

Nucleotide-dependent Movement of the ε Subunit between α and β Subunits in the Escherichia coli F1F0-type ATPase

Alanine-scanning Mutagenesis of the ∊ Subunit of the F1-F0 ATP Synthase from Escherichia coli Reveals Two Classes of Mutants

Contact Info

Product

Resources

About