Protocol for direct measurement of stability and activity of mitochondria electron transport chain complex II.

Electron Transport Complex II Electron Transport Biological Assay Cell Line Mitochondria
Cell Isolation Cell-based Assays Metabolism Protein Biochemistry

Journal

STAR protocols
ISSN: 2666-1667
Titre abrégé: STAR Protoc
Pays: United States
ID NLM: 101769501

Informations de publication

Date de publication:
17 03 2023
Historique:
received: 11 09 2022
revised: 06 11 2022
accepted: 15 12 2022
medline: 23 10 2023
pubmed: 10 1 2023
entrez: 9 1 2023
Statut: ppublish

Résumé

Mitochondria electron transport chain (ETC) complex II is essential for steroid metabolism. Here, we present a protocol to measure the stability and activity of mitochondria ETC complex II. We first describe mitochondria isolation from cell lines and tissues. We then detail how to determine the stability of ETC complex II using isothermal calorimetry and quantification of steroidogenesis using activity assays in parallel. Finally, we describe the steps to perform radioimmunoassay (RIA) to confirm the activity of ETC complex II. For complete details on the use and execution of this protocol, please refer to Bose et al. (2020).

Identifiants

DOI: 10.1016/j.xpro.2022.101996 PMID: 36620990 PMC: PMC9841272
pubmed: 36620990
pii: S2666-1667(22)00876-0
doi: 10.1016/j.xpro.2022.101996
pmc: PMC9841272
pii:
doi:

Substances chimiques

Electron Transport Complex II EC 1.3.5.1

Types de publication

Journal Article Research Support, N.I.H., Extramural Research Support, Non-U.S. Gov't

Langues

eng

Sous-ensembles de citation

IM

Pagination

101996

Informations de copyright

Copyright © 2022 The Author(s). Published by Elsevier Inc. All rights reserved.

Déclaration de conflit d'intérêts

Declaration of interests The authors declare no competing interests.

Auteurs

Himangshu S Bose (HS)

Laboratory of Biochemistry and Cell Biology, Department of Biomedical Sciences, Mercer University School of Medicine, Hoskins Research Building 1250 East 66th Street, Savannah, GA 31404, USA; Anderson Cancer Institute, Memorial University Medical Center, Savannah, GA 31404, USA. Electronic address: bose_hs@mercer.edu.

Nicole E Doetch (NE)

Laboratory of Biochemistry and Cell Biology, Department of Biomedical Sciences, Mercer University School of Medicine, Hoskins Research Building 1250 East 66th Street, Savannah, GA 31404, USA.

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Classifications MeSH

questionsmedicales.fr › Acides aminés, peptides et protéines › Protéines › Métalloprotéines › Protéines de liaison au fer › Protéines à fer non hème › Ferrosulfoprotéines › Complexe II de la chaîne respiratoire › Protocol for direct measurement of stability...
questionsmedicales.fr › Métabolisme › Voies et réseaux métaboliques › Transport d'électrons › Protocol for direct measurement of stability...
questionsmedicales.fr › Techniques d'investigation › Dosage biologique › Protocol for direct measurement of stability...
questionsmedicales.fr › Cellules › Cellules cultivées › Lignée cellulaire › Protocol for direct measurement of stability...
questionsmedicales.fr › Cellules › Structures cellulaires › Fractions subcellulaires › Mitochondries › Protocol for direct measurement of stability...