Table II Use of dodecylmaltoside-solubilized bovine heart mitochondria for mass calibration on two-dimensional BN/SDS gel

Dodecylmaltoside-solubilized RCs from bovine heart mitochondria were separated by BNE on a linear 3.5–16% acrylamide gradient gel followed by two-dimensional Tricine-SDS-PAGE using a 10% acrylamide gel. Mass/migration data refer to Fig. 3. Protein numbers 1–16 were identified by mass spectrometry as detailed in the supplemental material. Molecular weight search (MOWSE) scores larger than 65 were considered significant (p < 0.05) for protein identification.

SpotIdentified proteinUniProt accession no.MassapIaMigrationTMH per monomerGRAVY scoreMOWSE score
bcdef
kDamm
IRC I∼1000 (mon)<724.5
MMonomeric complex V597 (mon)<731.0
IIIRC III482 (dim)<737.5
IVRC IV205 (mon)<753.0
IIRC II123 (mon)<761.0
1aHemoglobin α chainP0196615.1 (mon)8.291.00000+0.01497
1bHemoglobin β chainP0207016.0 (mon)7.091.00000−0.004104
2MyoglobinP0219216.9 (mon)7.091.00000−0.37883
3Not identified91.0
4Rieske iron-sulfur protein of RC IIIP1327229.5 (mon)9.087.021001−0.125208
5AlbuminP0276966.4 (mon)5.679.50000−0.475117
675-kDa glucose-regulated proteinQ3ZCH068.7 (mon)5.571.51100−0.398183
7mAconitase 2, mitochondrial, monomerP2000482.4 (mon)7.268.51–2300−0.353330
7dAconitase 2, mitochondrial, dimerP20004164.8 (dim)7.258.01–2300−0.353137
8Nicotinamide nucleotide transhydrogenaseP11024109.0 (mon)7.360.51417121214+0.307413
9Succinate dehydrogenase, subunit AP31039123 (in RC II)6.461.01–2000−0.283242
103-Oxoacid CoA-transferase 1Q24JZ7113 (dim)8.662.03–4300−0.099128
11Citrate synthase, mitochondrialQ29RK198.0 (dim)7.071.52000−0.228156
12Isocitrate dehydrogenase 2, mitochondrialQ0446793.0 (dim)8.567.01000−0.382148
13Creatine kinase M chainQ9XSC686.0 (dim)6.665.00000−0.573110
14Malate dehydrogenase 2, mitochondrialQ32LG366.4 (dim)8.470.52–3000+0.126187
15Adenine nucleotide translocator 2Q8SQH565.6 (dim)9.874.53–44026+0.032119
16aSuccinate dehydrogenase, subunit BQ3T189123 (in RC II)8.561.01000−0.49196
16bEnoyl-CoA hydratase, short chain 1, mitochondrialQ58DM8168.6 (hex)6.961.01100+0.027358
  • a Calculated for the mature protein using the Swiss-Prot/UniProt database and the pI/Mw tool from the ExPASy proteomics server (www.expasy.ch). Information on the oligomeric state of the identified proteins was also taken from the Swiss-Prot/UniProt database. mon, monomer; dim, dimer; hex, hexamer.

  • b Content of TMHs analyzed by TMpred prediction program.

  • c Content of TMHs analyzed by HMMTOP prediction program.

  • d Content of TMHs analyzed by SOSUI prediction program.

  • e Content of TMHs analyzed by TMHMM prediction program.

  • f Experimental data for the Rieske iron-sulfur protein, adenine nucleotide translocator 2, and nicotinamide nucleotide transhydrogenase were taken from Refs. 10, 11, and 12, respectively.