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. 1971 Sep;68(9):2145–2148. doi: 10.1073/pnas.68.9.2145

Purification of the araC Protein

Gary Wilcox *, Kenneth J Clemetson †, Daniel V Santi †, Ellis Englesberg *
PMCID: PMC389371  PMID: 4943786

Abstract

The araC gene product, a regulatory protein required for expression of the L-arabinose operon, has been purified by affinity chromatography on Sepharose 4B to which 4-aminophenyl-β-D-6-deoxygalactopyranoside (an anti-inducer of the L-arabinose operon) had been covalently attached by means of a 4-aminophenylbutanamido side chain. Evidence is presented showing that the protein binds specifically to ara DNA.

Keywords: affinity chromatography, positive regulator, anti-inducer, ara operon

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Selected References

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

  1. Beverin S., Sheppard D. E., Park S. S. D-Fucose as a gratuitous inducer of the L-arabinose operon in strains of Escherichia coli B-r mutant in gene araC. J Bacteriol. 1971 Jul;107(1):79–86. doi: 10.1128/jb.107.1.79-86.1971. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Cuatrecasas P., Wilchek M., Anfinsen C. B. Selective enzyme purification by affinity chromatography. Proc Natl Acad Sci U S A. 1968 Oct;61(2):636–643. doi: 10.1073/pnas.61.2.636. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Emmer M., deCrombrugghe B., Pastan I., Perlman R. Cyclic AMP receptor protein of E. coli: its role in the synthesis of inducible enzymes. Proc Natl Acad Sci U S A. 1970 Jun;66(2):480–487. doi: 10.1073/pnas.66.2.480. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Englesberg E., Irr J., Power J., Lee N. Positive control of enzyme synthesis by gene C in the L-arabinose system. J Bacteriol. 1965 Oct;90(4):946–957. doi: 10.1128/jb.90.4.946-957.1965. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Englesberg E., Sheppard D., Squires C., Meronk F., Jr An analysis of "revertants" of a deletion mutant in the C gene of the L-arabinose gene complex in Escherichia coli B-r: isolation of initiator constitutive mutants (Ic). J Mol Biol. 1969 Jul 28;43(2):281–298. doi: 10.1016/0022-2836(69)90268-x. [DOI] [PubMed] [Google Scholar]
  6. Englesberg E., Squires C., Meronk F., Jr The L-arabinose operon in Escherichia coli B-r: a genetic demonstration of two functional states of the product of a regulator gene. Proc Natl Acad Sci U S A. 1969 Apr;62(4):1100–1107. doi: 10.1073/pnas.62.4.1100. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Hogg R. W., Englesberg E. L-arabinose binding protein from Escherichia coli B-r. J Bacteriol. 1969 Oct;100(1):423–432. doi: 10.1128/jb.100.1.423-432.1969. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Irr J., Englesberg E. Nonsense mutants in the regulator gene araC of the L-arabinose system of Escherichia coli B-r. Genetics. 1970 May;65(1):27–39. doi: 10.1093/genetics/65.1.27. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Katz L., Englesberg E. Hyperinducibility as a result of mutation in structural genes and self-catabolite repression in the ara operon. J Bacteriol. 1971 Jul;107(1):34–52. doi: 10.1128/jb.107.1.34-52.1971. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Kessler D. P., Englesberg E. Arabinose-leucine deletion mutants of Escherichia coli B-r. J Bacteriol. 1969 Jun;98(3):1159–1169. doi: 10.1128/jb.98.3.1159-1169.1969. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Kessler D. P., Englesberg E. Arabinose-leucine deletion mutants of Escherichia coli B-r. J Bacteriol. 1969 Jun;98(3):1159–1169. doi: 10.1128/jb.98.3.1159-1169.1969. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Porath J., Axen R., Ernback S. Chemical coupling of proteins to agarose. Nature. 1967 Sep 30;215(5109):1491–1492. doi: 10.1038/2151491a0. [DOI] [PubMed] [Google Scholar]
  13. Power J. The L-rhamnose genetic system in Escherichia coli K-12. Genetics. 1967 Mar;55(3):557–568. doi: 10.1093/genetics/55.3.557. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Riggs A. D., Suzuki H., Bourgeois S. Lac repressor-operator interaction. I. Equilibrium studies. J Mol Biol. 1970 Feb 28;48(1):67–83. doi: 10.1016/0022-2836(70)90219-6. [DOI] [PubMed] [Google Scholar]
  15. Schwartz M. Expression phénotypique et localisation génétique de mutations affectant le métabolisme du maltose chez Escherichia coli K 12. Ann Inst Pasteur (Paris) 1967 Jun;112(6):673–698. [PubMed] [Google Scholar]
  16. Sheppard D. E., Englesberg E. Further evidence for positive control of the L-arabinose system by gene araC. J Mol Biol. 1967 May 14;25(3):443–454. doi: 10.1016/0022-2836(67)90197-0. [DOI] [PubMed] [Google Scholar]
  17. Sheppard D., Englesberg E. Positive control in the L-arabinose gene-enzyme complex of Escherichia coli B/r exhibited with stable merodiploids. Cold Spring Harb Symp Quant Biol. 1966;31:345–347. doi: 10.1101/sqb.1966.031.01.044. [DOI] [PubMed] [Google Scholar]
  18. Steers E., Jr, Cuatrecasas P., Pollard H. B. The purification of beta-galactosidase from Escherichia coli by affinity chromatography. J Biol Chem. 1971 Jan 10;246(1):196–200. [PubMed] [Google Scholar]
  19. WILLSON C., PERRIN D., COHN M., JACOB F., MONOD J. NON-INDUCIBLE MUTANTS OF THE REGULATOR GENE IN THE "LACTOSE" SYSTEM OF ESCHERICHIA COLI. J Mol Biol. 1964 Apr;8:582–592. doi: 10.1016/s0022-2836(64)80013-9. [DOI] [PubMed] [Google Scholar]
  20. Zubay G., Schwartz D., Beckwith J. Mechanism of activation of catabolite-sensitive genes: a positive control system. Proc Natl Acad Sci U S A. 1970 May;66(1):104–110. doi: 10.1073/pnas.66.1.104. [DOI] [PMC free article] [PubMed] [Google Scholar]

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