The DOI matches its Crossref record; 12 of the 12 references it cites were found in a registry or an index.
Spectrophotometric Assay for Complex I of the Respiratory Chain in Tissue Samples and Cultured Fibroblasts
Abstract
Background: A reliable and sensitive complex I assay is an essential tool for the diagnosis of mitochondrial disorders, but current spectrophotometric assays suffer from low sensitivity, low specificity, or both. This deficiency is mainly due to the poor solubility of coenzyme-Q analogs and reaction mixture turbidity caused by the relatively high concentrations of tissue extract that are often required to measure complex I. Methods: We developed a new spectrophotometric assay to measure complex I in mitochondrial fractions and applied it to muscle and cultured fibroblasts. The method is based on measuring 2,6-dichloroindophenol reduction by electrons accepted from decylubiquinol, reduced after oxidation of NADH by complex I. The assay thus is designed to avoid nonspecific NADH oxidation because electrons produced in these reactions are not accepted by decylubiquinone, resulting in high rotenone sensitivity. Results: The assay was linear with time and amount of mitochondria. The Km values for NADH and 2,6-dichloroindophenol in muscle mitochondria were 0.04 and 0.017 mmol/L, respectively. The highest complex I activities were measured with 0.07 mmol/L decylubiquinone and 3.5 g/L bovine serum albumin. The latter was an essential component of the reaction mixture, increasing the solubility of decylubiquinone and rotenone. In patients with previously diagnosed complex I deficiencies, the new assay detected the complex I deficiencies in both muscle and fibroblasts. Conclusions: This spectrophotometric assay is reproducible, sensitive, and specific for complex I activity because of its high rotenone sensitivity, and it can be applied successfully to the diagnosis of complex I deficiencies.
Overview
The claims and statistics of this paper haven’t been checked yet.
Run the check — free, no account needed. It reads the open-access text and checks the sources it cites.
Integrity signals
Registry
Cites
Statistics
Verification
Claims the paper makes
The claims and statistics of this paper haven’t been checked yet.
Run the check — free, no account needed. It reads the open-access text and checks the sources it cites.
Self-check
Percentages, sums and reported differences re-computed from the paper’s own numbers.
The self-check needs the paper’s full text.
Citations
Cites
PROTEIN MEASUREMENT WITH THE FOLIN PHENOL REAGENT
Lowry, O., Rosebrough, N., Farr, A. L., & Randall, R. (1951) · Journal of Biological Chemistry · doi:10.1016/s0021-9258(19)52451-6 · Cited by 317,331
A MICROSPECTROPHOTOMETRIC METHOD FOR THE DETERMINATION OF CYTOCHROME OXIDASE
Cooperstein, S. J., & Lazarow, A. (1951) · Journal of Biological Chemistry · doi:10.1016/s0021-9258(18)44883-1 · Cited by 1,352
Biochemical and molecular investigations in respiratory chain deficiencies
Rustin, P., Chretien, D., Bourgeron, T., Gérard, B., Rötig, A., Saudubray, J. M., & Munnich, A. (1994) · Clinica Chimica Acta · doi:10.1016/0009-8981(94)90055-8 · Cited by 1,243
Bovine Complex I Is a Complex of 45 Different Subunits
Carroll, J., Fearnley, I. M., Skehel, J. M., Shannon, R. J., Hirst, J., & Walker, J. E. (2006) · Journal of Biological Chemistry · doi:10.1074/jbc.m607135200 · Cited by 487
Rotenone Model of Parkinson Disease
Panov, A., Dikalov, S., Shalbuyeva, N., Taylor, G., Sherer, T., & Greenamyre, J. T. (2005) · Journal of Biological Chemistry · doi:10.1074/jbc.m508628200 · Cited by 288
Mitochondrial complex I: structure, function and pathology
Janssen, R. J. R. J., Nijtmans, L. G., Heuvel, L. P. V. D., & Smeitink, J. A. M. (2006) · Journal of Inherited Metabolic Disease · doi:10.1007/s10545-006-0362-4 · Cited by 276
Self-citationFletcher, J. E., Spector, A. A., & Ashbrook, J. D. (1971) · Biochemistry · doi:10.1021/bi00793a011 · Cited by 269
Janssen, A. J. M., Trijbels, F. J. M., Sengers, R. C. A., Wintjes, L. T., Ruitenbeek, W., Smeitink, J. A. M., Morava, É., van Engelen, B. G. M., van den Heuvel, L. P., & Rodenburg, R. J. (2006) · Clinical Chemistry · doi:10.1373/clinchem.2005.062414 · Cited by 97
Self-citationEstimation of NADH oxidation in human skeletal muscle mitochondria
Fischer, J., Ruitenbeek, W., Trijbels, J. M. F., et al. (1986) · Clinica Chimica Acta · doi:10.1016/0009-8981(86)90246-9 · Cited by 85
Kramer, K. A., Oglesbee, D., Hartman, S. J., et al. (2005) · Clinical Chemistry · doi:10.1373/clinchem.2005.050146 · Cited by 60
Cited by
Lethal Cardiomyopathy in Mice Lacking Transferrin Receptor in the Heart
Xu, W., Barrientos, T., Mao, L., Rockman, H. A., Sauve, A. A., & Andrews, N. C. (2015) · Cell Reports · doi:10.1016/j.celrep.2015.09.023 · Cited by 275
Ibebunjo, C., Chick, J. M., Kendall, T., Eash, J. K., Li, C., Zhang, Y., Vickers, C., Wu, Z., Clarke, B. A., Shi, J., et al. (2013) · Molecular and Cellular Biology · doi:10.1128/mcb.01036-12 · Cited by 267
Role of sirtuins in lifespan regulation is linked to methylation of nicotinamide
Schmeisser, K., Mansfeld, J., Kuhlow, D., Weimer, S., Priebe, S., Heiland, I., Birringer, M., Groth, M., Segref, A., Kanfi, Y., et al. (2013) · Nature Chemical Biology · doi:10.1038/nchembio.1352 · Cited by 236
Acyl-CoA Dehydrogenase 9 Is Required for the Biogenesis of Oxidative Phosphorylation Complex I
Nouws, J., Nijtmans, L., Houten, S. M., van den Brand, M., Huynen, M., Venselaar, H., Hoefs, S., Gloerich, J., Kronick, J., Hutchin, T., et al. (2010) · Cell Metabolism · doi:10.1016/j.cmet.2010.08.002 · Cited by 190
Self-citationSangineto, M., Ciarnelli, M., Cassano, T., Radesco, A., Moola, A., Bukke, V. N., Romano, A., Villani, R., Kanwal, H., Capitanio, N., et al. (2023) · Redox Biology · doi:10.1016/j.redox.2023.102846 · Cited by 142
Metadata
- Authors
- Janssen, A. J., Trijbels, F. J., Sengers, R. C., Smeitink, J. A., van den Heuvel, L. P., Wintjes, L. T., Stoltenborg-Hogenkamp, B. J., & Rodenburg, R. J.
- Venue
- Clinical Chemistry
- Publisher
- Oxford University Press (OUP)
- Published
- 2007-04-01
- Volume
- 53
- Issue
- 4
- Pages
- 729-734
- ISSN
- 0009-9147 (ISSN-L)
- 1530-8561
- OpenAlex
- W2157999222
- Cited by
- 373
- References
- 12
- FWCI
- 5.75
- Type
- journal article
- Language
- en
- Open access
- closed
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
How to cite
Cite CoChat for the checks on this page, and the paper for its own findings.
- This page
- CoChat. (2026). Citation check: Spectrophotometric Assay for Complex I of the Respiratory Chain in Tissue Samples and Cultured Fibroblasts. https://app.cochat.ai/papers/10.1373/clinchem.2006.078873
- The paper
- Janssen, A. J., Trijbels, F. J., Sengers, R. C., Smeitink, J. A., van den Heuvel, L. P., Wintjes, L. T., Stoltenborg-Hogenkamp, B. J., & Rodenburg, R. J. (2007). Spectrophotometric Assay for Complex I of the Respiratory Chain in Tissue Samples and Cultured Fibroblasts. Clinical Chemistry. https://doi.org/10.1373/clinchem.2006.078873
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.
