Mitochondria isolated in nearly isotonic KCl buffer: Focus on cardiolipin and organelle morphology
Rat liver mitochondria were isolated in parallel in two different isolation buffers: a standard buffer containing mannitol/sucrose and a nearly physiological KCl based solution. The two different organelle preparations were comparatively characterized by respiratory activity, heme content, microsoma...
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| Published in | Biochimica et biophysica acta Vol. 1798; no. 3; pp. 681 - 687 |
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| Main Authors | , , , , , , |
| Format | Journal Article |
| Language | English |
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Elsevier B.V
01.03.2010
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| Abstract | Rat liver mitochondria were isolated in parallel in two different isolation buffers: a standard buffer containing mannitol/sucrose and a nearly physiological KCl based solution. The two different organelle preparations were comparatively characterized by respiratory activity, heme content, microsomal and Golgi contamination, electron microscopy and lipid analyses. The substitution of saccharides with KCl in the isolation buffer does not induce the formation of mitoplasts or disruption of mitochondria. Mitochondria isolated in KCl buffer are coupled and able to maintain a stable transmembrane charge separation. A number of biochemical and functional differences between the two organelle preparations are described; in particular KCl mitochondria exhibit lower cardiolipin content and smaller intracristal compartments in comparison with the standard mitochondrial preparation. |
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| AbstractList | Rat liver mitochondria were isolated in parallel in two different isolation buffers: a standard buffer containing mannitol/sucrose and a nearly physiological KCl based solution. The two different organelle preparations were comparatively characterized by respiratory activity, heme content, microsomal and Golgi contamination, electron microscopy and lipid analyses. The substitution of saccharides with KCl in the isolation buffer does not induce the formation of mitoplasts or disruption of mitochondria. Mitochondria isolated in KCl buffer are coupled and able to maintain a stable transmembrane charge separation. A number of biochemical and functional differences between the two organelle preparations are described; in particular KCl mitochondria exhibit lower cardiolipin content and smaller intracristal compartments in comparison with the standard mitochondrial preparation. |
| Author | Mastrodonato, Maria Lorusso, Michele Zaccagnino, Patrizia Liquori, Giuseppa E. Corcelli, Angela Lopalco, Patrizia Saponetti, Matilde Sublimi |
| Author_xml | – sequence: 1 givenname: Angela surname: Corcelli fullname: Corcelli, Angela email: a.corcelli@biologia.uniba.it organization: Department of Medical Biochemistry, Biology and Physics, University of Bari, Bari, Italy – sequence: 2 givenname: Matilde Sublimi surname: Saponetti fullname: Saponetti, Matilde Sublimi organization: Department of Medical Biochemistry, Biology and Physics, University of Bari, Bari, Italy – sequence: 3 givenname: Patrizia surname: Zaccagnino fullname: Zaccagnino, Patrizia organization: Department of Medical Biochemistry, Biology and Physics, University of Bari, Bari, Italy – sequence: 4 givenname: Patrizia surname: Lopalco fullname: Lopalco, Patrizia organization: Department of Medical Biochemistry, Biology and Physics, University of Bari, Bari, Italy – sequence: 5 givenname: Maria surname: Mastrodonato fullname: Mastrodonato, Maria organization: Department of Animal and Environmental Biology, University of Bari, Italy – sequence: 6 givenname: Giuseppa E. surname: Liquori fullname: Liquori, Giuseppa E. organization: Department of Animal and Environmental Biology, University of Bari, Italy – sequence: 7 givenname: Michele surname: Lorusso fullname: Lorusso, Michele organization: Department of Medical Biochemistry, Biology and Physics, University of Bari, Bari, Italy |
| BackLink | https://www.ncbi.nlm.nih.gov/pubmed/20093104$$D View this record in MEDLINE/PubMed |
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| Cites_doi | 10.1016/j.biocel.2009.04.010 10.1016/S0021-9258(19)39106-9 10.1074/jbc.273.31.19612 10.1038/sj.emboj.7600798 10.1021/bi048802k 10.1016/j.bbamem.2009.06.010 10.1093/oxfordjournals.jbchem.a135426 10.1074/jbc.M302021200 10.1038/nprot.2006.478 10.1006/bbrc.2000.3653 10.1139/o59-099 10.1074/jbc.M402545200 10.1038/sj.emboj.7601618 10.1111/j.1582-4934.2008.00281.x 10.1016/j.bbamcr.2006.04.006 10.1083/jcb.17.1.208 10.1194/jlr.M800312-JLR200 10.1016/S0163-7827(00)00005-9 10.1083/jcb.30.2.269 10.1016/S0021-9258(19)52749-1 10.1016/S0021-9258(19)43371-1 10.1083/jcb.37.2.345 10.1074/jbc.M202396200 10.1529/biophysj.107.126920 |
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| Keywords | Mitochondrion KCl based isolation buffer Mitochondrial lipids Cardiolipin Cristae morphology |
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| SubjectTerms | Animals Buffers Cardiolipin Cardiolipins - metabolism Cell Respiration - drug effects Chromatography, Thin Layer Cristae morphology Cytochrome Reductases - metabolism Heme - metabolism Isotonic Solutions - pharmacology KCl based isolation buffer Lipids - analysis Mitochondria, Liver - drug effects Mitochondria, Liver - enzymology Mitochondria, Liver - metabolism Mitochondria, Liver - ultrastructure Mitochondrial lipids Mitochondrion NADP - metabolism Organelle Shape - drug effects Oxygen Consumption - drug effects Potassium Chloride - pharmacology Rats Spectrometry, Mass, Electrospray Ionization |
| Title | Mitochondria isolated in nearly isotonic KCl buffer: Focus on cardiolipin and organelle morphology |
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