Subunits of succinyl-coenzyme A synthetase: coordination of production in Escherichia coli and discovery of a factor that precludes refolding

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Abstract
Succinyl-coenzyme A synthetase of Escherichia coli has an alpha 2 beta 2 subunit structure. By measuring reconstituted enzyme activity present after addition of purified alpha or beta subunits to cell extracts followed by refolding, we have shown that extracts contain no significant excess of either subunit species. This equivalence suggests that the expression of the respective structural genes for the subunits is coordinately controlled. The presence of cell extract does not affect the rate or extent of reassembly of the subunits, pointing to a high degree of specificity of mutual recognition by the refolding subunits. In the course of these experiments, we have detected the presence in cell extracts of a low-molecular-weight factor that specifically inactivates unfolded alpha or beta subunits or prevents their reassembly into catalytically active enzyme. Under conditions where the subunits are completely inactivated, the factor has no detectable effect on native or refolded tetrameric enzyme, suggesting that the factor may react only with unfolded protein.