The DOI matches its Crossref record; 42 of the 42 references it cites were found in a registry or an index.
A Functionally Active Human F1F0 ATPase Can be Purified by Immunocapture from Heart Tissue and Fibroblast Cell Lines
Open accessAbstract
Human mitochondrial F(1)F(0) ATP synthase was isolated with a one-step immunological approach, using a monoclonal antibody against F(1) in a 96-well microplate activity assay system, to establish a method for fast high throughput screening of inhibitors, toxins, and drugs with very small amounts of enzyme. For preparative purification, mitochondria from human heart tissue as well as cultured fibroblasts were solubilized with dodecyl-beta-d-maltoside, and the F(1)F(0) was isolated with anti-F(1) monoclonal antibody coupled to protein G-agarose beads. The immunoprecipitated F(1)F(0) contained a full complement of subunits that were identified with specific antibodies against five of the subunits (alpha, beta, OSCP, d, and IF(1)) and by MALDI-TOF and/or LC/MS/MS for all subunits except subunit c, which could not be resolved by these methods because of the limits of detection. Microscale immunocapture of F(1)F(0) from detergent-solubilized mitochondria or whole cell fibroblast extracts was performed using anti-F(1) monoclonal antibody immobilized on 96-well microplates. The captured complex V displayed ATP hydrolysis activity that was fully oligomycin and inhibitor protein IF(1)-sensitive. Moreover, IF(1) could be co-isolated with F(1)F(0) when the immunocapture procedure was carried out at pH 6.5 but was absent when the ATP synthase was isolated at pH 8.0. Immunocaptured F(1)F(0) lacking IF(1) could be inhibited by more than 90% by addition of recombinant inhibitor protein, and conversely, F(1)F(0) containing IF(1) could be activated more than 10-fold by brief exposure to pH 8.0, inducing the release of inhibitor protein. With this microplate system an ATP hydrolysis assay of complex V could be carried out with as little as 10 ng of heart mitochondria/well and as few as 3 x 10(4) cells/well from fibroblast cultures. The system is therefore suitable to screen patient-derived samples for alterations in amount or functionality of both the F(1)F(0) ATPase and IF(1).
Overview
Integrity signals
Registry
Cites
Statistics
Verification
Claims the paper makes
Claims other papers make about this one
Each is checked against this paper’s own text: does it say what the citing paper says it does?
In the citing paper
“These diseases’ causal genes co-localize with ATP5PO62–65.”
From Few shot learning for phenotype-driven diagnosis of patients with rare genetic diseases (2025) · cites this paper as [63]
Citations
Cites
Cleavage of Structural Proteins during the Assembly of the Head of Bacteriophage T4
LAEMMLI, U. K. (1970) · Nature · doi:10.1038/227680a0 · Cited by 249,961
THE ATP SYNTHASE—A SPLENDID MOLECULAR MACHINE
Boyer, P. D. (1997) · Annual Review of Biochemistry · doi:10.1146/annurev.biochem.66.1.717 · Cited by 2,014
Supercomplexes in the respiratory chains of yeast and mammalian mitochondria
Schägger, H., & Pfeiffer, K. (2000) · The EMBO Journal · doi:10.1093/emboj/19.8.1777 · Cited by 1,356
Clauser, K. R., Baker, P. R., & Burlingame, A. L. (1999) · Analytical Chemistry · doi:10.1021/ac9810516 · Cited by 1,084
A one-step purification of membrane proteins using a high efficiency immunomatrix.
Schneider, C., Newman, R., Sutherland, D. R., Asser, U., & Greaves, M. F. (1982) · Journal of Biological Chemistry · doi:10.1016/s0021-9258(18)33889-4 · Cited by 764
Tatuch, Y., Christodoulou, J., Feigenbaum, A., Clarke, J. T. R., Wherret, J., Smith, C., Rudd, N., Petrova-Benedict, R., & Robinson, B. H. (1992) · PubMed · Cited by 490
Hu neuronal proteins are expressed in proliferating neurogenic cells
Marusich, M. F., Furneaux, H. M., Henion, P. D., & Weston, J. A. (1994) · Journal of Neurobiology · doi:10.1002/neu.480250206 · Cited by 486
Self-citationMitochondrial DNA Mutations and Pathogenesis
Schon, E. A., Bonilla, E., & DiMauro, S. (1997) · Journal of Bioenergetics and Biomembranes · doi:10.1023/a:1022685929755 · Cited by 424
Mitochondrial DNA mutations in human disease
DiMauro, S., & Schon, E. A. (2001) · American Journal of Medical Genetics · doi:10.1002/ajmg.1392 · Cited by 417
Primary structure and subunit stoichiometry of F1-ATPase from bovine mitochondria.
Walker, J., Fearnley, I., Gay, N. J., Gibson, B., Northrop, F., Powell, S., Runswick, M., Saraste, M., & Tybulewicz, V. (1985) · Journal of Molecular Biology · doi:10.1016/0022-2836(85)90313-4 · Cited by 392
Catalytic mechanism of F1-ATPase
Weber, J., & Senior, A. E. (1997) · Biochimica et Biophysica Acta (BBA) - Bioenergetics · doi:10.1016/s0005-2728(96)00121-1 · Cited by 378
Detailed peptide characterization using PEPTIDEMASS – a World‐Wide‐Web‐accessible tool
Wilkins, M. R., Lindskog, I., Gasteiger, E., Bairoch, A., Sanchez, J., Hochstrasser, D. F., & Appel, R. D. (1997) · Electrophoresis · doi:10.1002/elps.1150180314 · Cited by 358
Berggren, K. N., Chernokalskaya, E., Steinberg, T. H., Kemper, C., Lopez, M. F., Diwu, Z., Haugland, R. P., & Patton, W. F. (2000) · Electrophoresis · doi:10.1002/1522-2683(20000701)21:12 · Cited by 357
Mechanism of the F1F0-type ATP synthase, a biological rotary motor
Capaldi, R., & Aggeler, R. J. (2002) · Trends in Biochemical Sciences · doi:10.1016/s0968-0004(01)02051-5 · Cited by 335
Self-citationDucret, A., van Oostveen, I., Eng, J. K., Yates, J. R., & Aebersold, R. H. (1998) · Protein Science · doi:10.1002/pro.5560070320 · Cited by 310
Matsuyama, S., Xu, Q., Velours, J., & Reed, J. C. (1998) · Molecular Cell · doi:10.1016/s1097-2765(00)80033-7 · Cited by 270
Modulation of the Oligomerization State of the Bovine F1-ATPase Inhibitor Protein, IF1, by pH
Cabezón, E., Butler, P. J. G., Runswick, M. J., & Walker, J. E. (2000) · Journal of Biological Chemistry · doi:10.1074/jbc.m003859200 · Cited by 192
Collinson, I. R., Runswick, M. J., Buchanan, S. K., Fearnley, I. M., Skehel, M. J., van Raaij, M. J., Griffiths, D. E., & Walker, J. E. (1994) · Biochemistry · doi:10.1021/bi00191a026 · Cited by 167
Impaired ATP Synthase Assembly Associated with a Mutation in the Human ATP Synthase Subunit 6 Gene
Nijtmans, L. G. J., Henderson, N. S., Attardi, G. M., & Holt, I. J. (2001) · Journal of Biological Chemistry · doi:10.1074/jbc.m008114200 · Cited by 136
Regulation of the mitochondrial ATP synthase/ATPase complex
Schwerzmann, K., & Pedersen, P. L. (1986) · Archives of Biochemistry and Biophysics · doi:10.1016/0003-9861(86)90695-8 · Cited by 131
Garcı́a, J. J., Ogilvie, I. M., Robinson, B. H., & Capaldi, R. (2000) · Journal of Biological Chemistry · doi:10.1074/jbc.275.15.11075 · Cited by 122
Self-citationNovel features in the structure of bovine ATP synthase
Karrasch, S., & Walker, J. E. (1999) · Journal of Molecular Biology · doi:10.1006/jmbi.1999.2897 · Cited by 112
Marusich, M. F., Robinson, B. H., Taanman, J., Kim, S. J., Schillace, R. V., Smith, J. L., & Capaldi, R. (1997) · Biochimica et Biophysica Acta (BBA) - Molecular Basis of Disease · doi:10.1016/s0925-4439(97)00061-6 · Cited by 111
Self-citationThe regulation of catalysis in ATP synthase
Walker, J. E. (1994) · Current Opinion in Structural Biology · doi:10.1016/0959-440x(94)90274-7 · Cited by 104
The IF1 inhibitor protein of the mitochondrial F1F0-ATPase
Green, D. W., & Grover, G. J. (2000) · Biochimica et Biophysica Acta (BBA) - Bioenergetics · doi:10.1016/s0005-2728(00)00085-2 · Cited by 100
Mitochondrial Events in the Life and Death of Animal Cells: A Brief Overview
Pedersen, P. L. (1999) · Journal of Bioenergetics and Biomembranes · doi:10.1023/a:1005453700533 · Cited by 96
Wood, R. D., Biggerstaff, M., & Shivji, M. K. K. (1995) · Methods · doi:10.1006/meth.1995.1022 · Cited by 81
The ATPase Inhibitor Protein from Bovine Heart Mitochondria: The Minimal Inhibitory Sequence
van Raaij, M. J., Orriss, G. L., Montgomery, M. G., Runswick, M. J., Fearnley, I. M., Skehel, M. J., & Walker, J. E. (1996) · Biochemistry · doi:10.1021/bi960628f · Cited by 80
Large-scale chromatographic purification of F1F0-ATPase and complex I from bovine heart mitochondria
Buchanan, S. K., & Walker, J. E. (1996) · Biochemical Journal · doi:10.1042/bj3180343 · Cited by 70
Regulation of the mitochondrial ATPasein situ in cardiac muscle: Role of the inhibitor subunit
ROUSLIN, W. (1991) · Journal of Bioenergetics and Biomembranes · doi:10.1007/bf00786006 · Cited by 63
Triepels, R. H., Hanson, B. J., van den Heuvel, L. P., Sundell, L., Marusich, M. F., Smeitink, J. A. M., & Capaldi, R. (2001) · Journal of Biological Chemistry · doi:10.1074/jbc.m009903200 · Cited by 60
Self-citationInteraction between the mitochondrial ATP synthetase and ATPase inhibitor protein
Panchenko, M. V., & Vinogradov, A. D. (1985) · FEBS Letters · doi:10.1016/0014-5793(85)80611-6 · Cited by 58
Lutter, R., Saraste, M., WALRAVEN, H. S. V., Runswick, M. J., Finel, M., Deatherage, J. F., & Walker, J. E. (1993) · Biochemical Journal · doi:10.1042/bj2950799 · Cited by 44
Ko, Y. H., Pan, W., Inoue, C. N., & Pedersen, P. L. (2002) · Mitochondrion · doi:10.1016/s1567-7249(01)00036-8 · Cited by 42
Efficient hybridoma production using previously frozen splenocytes
Marusich, M. F. (1988) · Journal of Immunological Methods · doi:10.1016/0022-1759(88)90167-6 · Cited by 42
Self-citation[11] Structural analysis of ATP synthase from bovine heart mitochondria
Walker, J. E., Collinson, I. R., van Raaij, M. J., & Runswick, M. J. (1995) · Methods in enzymology on CD-ROM/Methods in enzymology · doi:10.1016/0076-6879(95)60136-8 · Cited by 41
Yamada, E. W., & Huzel, N. J. (1992) · Biochimica et Biophysica Acta (BBA) - Molecular Basis of Disease · doi:10.1016/0925-4439(92)90093-3 · Cited by 36
Factor B and the Mitochondrial ATP Synthase Complex
Belogrudov, G. I., & Hatefi, Y. (2002) · Journal of Biological Chemistry · doi:10.1074/jbc.m111256200 · Cited by 36
López-Mediavilla, C., Faure, H., & Godinot, C. (1993) · European Journal of Biochemistry · doi:10.1111/j.1432-1033.1993.tb18058.x · Cited by 31
Mitochondrial F0F1 ATP Synthase
Ko, Y. H., Hullihen, J. M., Hong, S., & Pedersen, P. L. (2000) · Journal of Biological Chemistry · doi:10.1074/jbc.m004453200 · Cited by 30
Zhang, X. H., Pan, W., & Hutchins, J. B. (1995) · Journal of Neurochemistry · doi:10.1046/j.1471-4159.1995.65062812.x · Cited by 22
Ziegler, M., & Penefsky, H. S. (1993) · Journal of Biological Chemistry · doi:10.1016/s0021-9258(19)74394-4 · Cited by 19
Research works printed but not linked: 2
- Tatuch, 1992, Am. J. Human Genet., 50, 852
- McGeoch, 2001, Eur. J. Pediatr. Neurol., 5A, 1447
Cited by
Characterization of the human heart mitochondrial proteome
Taylor, S. W., Fahy, E., Zhang, B., Glenn, G. M., Warnock, D. E., Wiley, S. E., Murphy, A. N., Gaucher, S. P., Capaldi, R., Gibson, B. W., et al. (2003) · Nature Biotechnology · doi:10.1038/nbt793 · Cited by 713
Self-citationYang, Z., Yan, C., Ma, J., Peng, P., Ren, X., Cai, S., Shen, X., Wu, Y. C., Zhang, S., Wang, X., et al. (2023) · Nature Metabolism · doi:10.1038/s42255-022-00710-w · Cited by 664
Schulenberg, B., Aggeler, R. J., Beechem, J., Capaldi, R., & Patton, W. F. (2003) · Journal of Biological Chemistry · doi:10.1074/jbc.c300189200 · Cited by 231
Self-citationXie, J., Marusich, M. F., Souda, P., Whitelegge, J. P., & Capaldi, R. (2007) · FEBS Letters · doi:10.1016/j.febslet.2007.06.052 · Cited by 226
Self-citationKo, Y. H., Delannoy, M. R., Hullihen, J. M., Chiu, W., & Pedersen, P. L. (2003) · Journal of Biological Chemistry · doi:10.1074/jbc.c200703200 · Cited by 192
Metadata
- Authors
- Aggeler, R., Coons, J., Taylor, S. W., Ghosh, S. S., Garcıáa, J. J., Capaldi, R. A., & Marusich, M. F.
- Affiliations
- University of Oregon (US)
- Instituto Nacional de Cardiología (MX)
- Corresponding author
- Capaldi, Roderick A.
- ORCID
- Ghosh, Soumitra S.
- Venue
- Journal of Biological Chemistry
- Publisher
- Elsevier BV
- Published
- 2002-09
- Volume
- 277
- Issue
- 37
- Pages
- 33906-33912
- ISSN
- 0021-9258 (ISSN-L)
- OpenAlex
- W2015413569
- Cited by
- 72
- References
- 42
- Citations by year
- 2026: 1 · 2025: 2 · 2024: 2 · 2023: 2 · 2022: 3 · 2020: 2 · 2018: 2 · 2017: 2 · 2016: 1 · 2015: 2 · 2014: 2 · 2013: 2 · 2012: 5
- FWCI
- 2.56
- Type
- journal article
- Language
- en
- Open access
- hybrid
- doi.org
- Licence
- Locations
- Journal of Biological Chemistry (publishedVersion, cc-by)
- PubMed (publishedVersion)
- Funders
- National Institutes of Health (5r01hl024526-25)
- National Heart, Lung, and Blood Institute (HL24526)
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › ATP Synthase and ATPases Research
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Mitochondrial Function and Pathology
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Structural Biology › Enzyme Structure and Function
- Keywords
- F0F1-ATP synthase, mitochondrial complex V, immunocapture, pH-dependent activation, high-throughput screening, fibroblasts, oligomycin sensitivity, subunit composition, drug screening, patient-derived samples
- MeSH terms
- Antibodies, Monoclonal, Cell Line, Fibroblasts, Proton-Translocating ATPases, Humans, Hydrogen-Ion Concentration, Myocardium, Precipitin Tests, Protein Subunits
About these checks
Registry, retraction, Cites and statistics results are computed, not judged, with one exception: a printed reference that no index lists is read once by a model before it counts as a citation error. Claims are extracted by a model and kept only when their quote appears verbatim in the text; each cited source is then checked, with a model’s help, for whether it says what the claim relies on. Those verdicts can be wrong — read the claim’s quote and the cited source before relying on one.
Statistics and arithmetic are recomputed, not judged.
Confidence bands are shown only for fields calibrated on our annotated test set. None is calibrated yet, so no band is shown.
What counts as a claim ›How we measure confidence ›How we check this ›
Are you an author, or do you see a problem with this page? Request a review
Data sources
Bibliographic data: Semantic Scholar, OpenAlex, Crossref.
Contains information from the Semantic Scholar Academic Graph Datasets, made available under the ODC Attribution License (ODC-BY 1.0).
Semantic Scholar’s data is described in: Kinney, R., et al. (2023). The Semantic Scholar Open Data Platform. arXiv:2301.10140.
