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. 1977 Oct;74(10):4223–4227. doi: 10.1073/pnas.74.10.4223

Acyl group and electron pair relay system: A network of interacting lipoyl moieties in the pyruvate and α-ketoglutarate dehydrogenase complexes from Escherichia coli

Jimmy H Collins 1,2, Lester J Reed 1,2
PMCID: PMC431911  PMID: 200908

Abstract

The dihydrolipoyl transacetylase component of the Escherichia coli pyruvate dehydrogenase complex [pyruvate:lipoate oxidoreductase (decarboxylating and acceptor-acetylating), EC 1.2.4.1] bears two sites on each of its 24 polypeptide chains that undergo reductive acetylation by [2-14C]pyruvate and thiamin pyrophosphate, acetylation by [1-14C]acetyl-CoA in the presence of DPNH, and reaction with N-ethyl[2,3-14C]maleimide in the presence of pyruvate and thiamin pyrophosphate. The data strongly imply that these sites are covalently bound lipoyl moieties. The results of similar experiments with the E. coli α-ketoglutarate dehydrogenase complex [2-oxoglutarate:lipoate oxidoreductase (decarboxylating and acceptor-succinylating), EC 1.2.4.2] indicate that its dihydrolipoyl transsuccinylase component bears only one lipoyl moiety on each of its 24 chains. Charging of the 48 acetyl acceptor sites on the transacetylase or the 24 succinyl acceptor sites on the transsuccinylase by pyruvate or α-ketoglutarate, respectively, and thiamin pyrophosphate was observed in the presence of only a few functionally active pyruvate dehydrogenase or α-ketoglutarate dehydrogenase chains. Extensive crosslinking of the transacetylase chains was observed when the pyruvate dehydrogenase complex was treated with pyruvate and thiamin pyrophosphate or with DPNH in the presence of N,N′-o- or N,N′-p-phenylenedimaleimide, respectively. When the α-ketoglutarate dehydrogenase complex was treated with DPNH in the presence of N,N′-p-phenylenedimaleimide, only transsuccinylase monomers and crosslinked transsuccinylase dimers were detected. It appears that the 48 lipoyl moieties in the transacetylase and the 24 lipoyl moieties in the transsuccinylase comprise an interacting network that functions as an acyl group and electron pair relay system through thiol-disulfide and acyl-transfer reactions among all of the lipoyl moieties.

Keywords: multienzyme complexes, thiol-disulfide interchange, crosslinking of subunits

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

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