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Assessment of Respi...
Assessment of Respiratory Enzymes in Intact Cells by Permeabilization with Alamethicin
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- Rasmusson, Allan G. (författare)
- Lund University,Lunds universitet,Molekylär cellbiologi,Biologiska institutionen,Naturvetenskapliga fakulteten,Växtbiologi,Forskargrupper vid Lunds universitet,Molecular Cell Biology,Department of Biology,Faculty of Science,Plant Biology,Lund University Research Groups
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- Møller, Ian Max (författare)
- Aarhus University
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- Widell, Susanne (författare)
- Lund University,Lunds universitet,Molekylär cellbiologi,Biologiska institutionen,Naturvetenskapliga fakulteten,Molecular Cell Biology,Department of Biology,Faculty of Science
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Van Aken, Olivier (redaktör/utgivare)
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Rasmusson, Allan g. (redaktör/utgivare)
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(creator_code:org_t)
- 2021-09-21
- 2022
- Engelska 8 s.
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Ingår i: Plant Mitochondria : Methods and Protocols - Methods and Protocols. - New York, NY : Springer US. - 1064-3745 .- 1940-6029. - 9781071616529 - 9781071616536 ; 2363, s. 77-84
- Relaterad länk:
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http://dx.doi.org/10...
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https://lup.lub.lu.s...
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https://doi.org/10.1...
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Abstract
Ämnesord
Stäng
- We here describe measurements of respiratory enzymes in situ, which can be done on very small cell samples and make mitochondrial isolation unnecessary. The method is based on the ability of the fungal peptide alamethicin to permeate biological membranes from the net positively charged side, and form nonspecific ion channels. These channels allow rapid transport of substrates and products across the plasma membrane, the inner mitochondrial membrane, and the inner plastid envelope. In this way, mitochondrial enzyme activities can be studied without disrupting the cells. The enzymes can be investigated in their natural proteinaceous environment and the activity of enzymes, also those sensitive to detergents or to dilution, can be quantified on a whole cell basis. We here present protocols for in situ measurement of two mitochondrial enzymatic activities: malate oxidation measured as oxygen consumption by the electron transport chain, which is sensitive to detergents, and NAD+-isocitrate dehydrogenase, a tricarboxylic acid cycle enzyme that dissociates upon dilution.
Ämnesord
- NATURVETENSKAP -- Biologi -- Biokemi och molekylärbiologi (hsv//swe)
- NATURAL SCIENCES -- Biological Sciences -- Biochemistry and Molecular Biology (hsv//eng)
Nyckelord
- Alamethicin
- BY-2 cells
- Electron transport chain
- Mitochondria
- Tricarboxylic acid cycle
Publikations- och innehållstyp
- kap (ämneskategori)
- ref (ämneskategori)
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