Purification of Tyrosine-Sensitive 3-Deoxy-D-Arabinoheptulosonate-7-Phosphate (DAHP) and Tyrosyl-tRNA Synthetases on Agarose Carrying Carboxyl-Linked Tyrosine

  • Andrew R. Gallopo
  • Philip S. Kotsiopoulos
  • Scott C. Mohr
Part of the Advances in Experimental Medicine and Biology book series (AEMB, volume 42)

Abstract

In principle any ligand-binding site with an appreciable degree of specificity can be used to purify a macromolecule by affinity chromatography. We have prepared a resin which contains covalently bound tyrosyl groups and employed it to purify one enzyme (tyrosyl-tRNA synthetase from E. coli) which interacts with the resin via its substrate-binding site and another (DAHP synthetase from yeast) which interacts via an effector-binding site. Our resin which contains a hexamethylene diamine (HMD) extension between the agarose matrix and the ligand appears to bind DAHP synthetase much more strongly than the previously used “tyrosineSepharose” which lacked such a spacer (1).

Keywords

Cellulose Tyrosine Agarose Macromolecule Phenylalanine 

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Copyright information

© Plenum Press, New York 1974

Authors and Affiliations

  • Andrew R. Gallopo
    • 1
  • Philip S. Kotsiopoulos
    • 2
  • Scott C. Mohr
    • 2
  1. 1.Department of ChemistryMontclair State CollegeUpper MontclairUSA
  2. 2.Department of ChemistryBoston UniversityBostonUSA

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