Association of calcineurin with mitochondrial proteins

Proteins
Abigail M Tokheim, Bruce L Martin

Abstract

Using mouse hearts from Swiss Webster mice, calcineurin was immunoprecipitated using commercially available anti-calcineurin antibody and the resulting complex analyzed by using sodium dodecyl sulfate-gel electrophoresis with silver staining. Distinct proteins were observed and subjected to in situ trypsin digestion followed by extraction of the resulting peptides. Peptides from each protein band were loaded onto a target for matrix-assisted laser desorption/ionization time-of-flight mass spectrometry and analyzed. The resulting peptide mass spectrum was compared with the Mascot and Protein Prospector databases and resulted in the specific identification of heart mitochondrial proteins, specifically Mn-superoxide dismutase (SOD), aconitase (ACN), and malate dehydrogenase (MDH). Each of the three mitochondrial enzymes was identified with approximately 15-25% sequence coverage and all with statistical significance (P < 0.05) according to the Mascot database search engine. Tandem mass spectrometry analysis of the peptide fragmentation spectra confirmed the identification of these protein partners and also yielded the identification of mitochondrial isocitrate dehydrogenase (ICDH) as another protein in the immunoprecipitated comple...Continue Reading

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Citations

Nov 4, 2008·Biochemistry. Biokhimii︠a︡·L V Matasova, T N Popova
Jun 8, 2012·International Journal of Cell Biology·Katarína Smolková, Petr Ježek
Apr 14, 2015·Nature Communications·Susan ChengThomas J Wang
Jul 7, 2007·American Journal of Physiology. Cell Physiology·Shi-Jin ZhangAbram Katz
Aug 30, 2020·Cell Communication and Signaling : CCS·Trevor P Creamer
Jan 15, 2008·American Journal of Physiology. Heart and Circulatory Physiology·Ilka PinzJoanne S Ingwall

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