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Synthesis of the Escherichia coli K12 isoenzymes of ornithine transcarbamylase, performed in vitro

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Summary

The in vitro synthesis of enzymaticallyactive ornithine transcarbamylase (OTCase) directed by each of the E. coli K-12 OTCase genes (argF and argI) is described. The E. coli OTCase isoenzyme subunits are not identical, whether synthesized in vivo or in vitro, the argF-coded product being about 5% smaller. The OTCase protomers are enzymatically inactive but associate in vitro to an enzymatically active multimer. The rates of subunit association of argF and argI isoenzymes are considerably different. Utilizing the facile assay protocol presented, the regulation of in vitro OTCase synthesis by the specific holorepressor of the arginine regulon is demonstrated. Calculations based upon data presented indicate that there are about 65 molecules of argR gene product per bacterium, a substantially lower estimate than previously reported.

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Author notes

  1. Michael L. Cleary & Eric James

    Present address: Department of Biochemistry, University of Kentucky, College of Medicine, 40506, Lexington, KY, USA

  2. Robert T. Garvin

    Present address: Department of Microbiology, Saint Louis University School of Medicine, 63104, St. Louis, MO, USA

Authors and Affiliations

  1. Department of Chemistry, University of South Carolina, 29208, Columbia, SC, USA

    Michael L. Cleary, Robert T. Garvin & Eric James

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  1. Michael L. Cleary
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  2. Robert T. Garvin
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  3. Eric James
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Additional information

Communicated by F. Gros

This work is dedicated to Luigi Gorini without whom none of this would have been possible. His unbounded love of science and freedom will be remembered by so many, for so long.

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Cleary, M.L., Garvin, R.T. & James, E. Synthesis of the Escherichia coli K12 isoenzymes of ornithine transcarbamylase, performed in vitro. Molec. Gen. Genet. 157, 155–165 (1977). https://doi.org/10.1007/BF00267393

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  • DOI: https://doi.org/10.1007/BF00267393

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