The purification and characterization of ATP synthase complexes from the mitochondria of four fungal species

March 19, 2015, MRC Mitochondrial Biology Unit

ATP, the fuel of life, is produced in mitochondria of living cells by a molecular machine, the ATP synthase. We have isolated the machines from four fungal species, compared their stabilities and identified the proteins from which they are constructed.

The ATP synthases have been isolated by affinity chromatography from the mitochondria of the , Yarrowia histolytica, Pichia pastoris, Pichia angusta and Saccharomyces cerevisae. The subunit compositions of the purified enzyme complexes depended upon the detergent used to solubilise and purify the complex, and the presence or absence of exogenous phospholipids.

All four enzymes purified in the presence of n-dodecyl-β-D-maltoside had a complete complement of core subunits involved directly in the synthesis of ATP, but they were deficient to different extents in their supernumerary membrane subunits. In contrast, the enzymes from P. angusta and S. cerevisiae purified in the presence of n-decyl-β-maltose neopentyl glycol and the phospholipids 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine, 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphoethanolamine, cardiolipin and 1-palmitoyl-2-oleoyl-sn-glycero-3-[phospho-rac-(1-glycerol)] had a complete complement of core subunits and also contained all the known supernumerary membrane subunits, e, f, g, j, k and ATP8 (or Aap1), plus an additional new membrane component named subunit l, related in sequence to subunit k.

The catalytic domain of the enzyme from P. angusta was more resistant to thermal denaturation than the enzyme from S. cerevisiae, but less stable than the of the bovine enzyme, but the stator and the integrity of the transmembrane proton pathway were most stable in the enzyme from P. angusta. The P. angusta enzyme provides a suitable source of for studying the structure of the membrane domain and properties associated with that sector of the .

Explore further: Transport molecule forms a protective structure to guide proteins to cell membrane

More information: "The purification and characterization of ATP synthase complexes from the mitochondria of four fungal species." Sidong Liu, Thomas J Charlesworth, John V Bason, Martin G Montgomery, Michael E Harbour, Ian M. Fearnley and John E Walker Biochem. J. (2015) DOI: 10.1042/BJ20150197

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Whydening Gyre
not rated yet Mar 22, 2015
A pretty dry and non laymen-type article -
nicely done...
Dethe
5 / 5 (1) Mar 22, 2015
These molecules should rotate or they're broken...

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