Cryoelectron microscopy of mammalian pyruvate dehydrogenase complex.
Cryoelectron microscopy has been performed on frozen-hydrated pyruvate dehydrogenase complexes from bovine heart and kidney and on various subcomplexes consisting of the dihydrolipoyl transacetylase-based (E2) core and substoichiometric levels of the other two major components, pyruvate dehydrogenase (E1) and dihydrolipoyl dehydrogenase (E3). The diameter of frozen-hydrated pyruvate dehydrogenase complex (PDC) is 50 nm, which is significantly larger than previously reported values. On the basis of micrographs of the subcomplexes, it is concluded that the E1 and E3 are attached to the E2-core complex by extended (4-6 nm maximally) flexible tethers. PDC constructed in this manner would probably collapse and appear smaller than its native size when dehydrated, as was the case in previous electron microscopy studies. The tether linking E1 to the core involves the hinge sequence located between the E1-binding and catalytic domains in the primary sequence of E2, whereas the tether linking E3 is probably derived from a similar hinge-type sequence in component X. Tilting of the E2-based cores and comparison with model structures confirmed that their overall shape is that of a pentagonal dodecahedron. The approximately 6 copies of protein X present in PDC do not appear to be clustered in one or two regions of the complex and are not likely to be symmetrically distributed.
Wagenknecht
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Wadsworth Center for Laboratories and Research, New York State Department of Health, Albany 12201-0509.
Grassucci
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Radke
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Roche
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D000123:Acetyltransferases / Q000378:metabolism; D000818:Animals; D002417:Cattle; D015925:Cryopreservation; D008058:Dihydrolipoamide Dehydrogenase / Q000378:metabolism; D051047:Dihydrolipoyllysine-Residue Acetyltransferase; D007668:Kidney / Q000201:enzymology*; D008854:Microscopy, Electron / Q000379:methods; D009206:Myocardium / Q000201:enzymology*; D011768:Pyruvate Dehydrogenase Complex / Q000378:metabolism / Q000648:ultrastructure*
D011768:Pyruvate Dehydrogenase Complex; D008058:Dihydrolipoamide Dehydrogenase; D000123:Acetyltransferases; D051047:Dihydrolipoyllysine-Residue Acetyltransferase