Purification of tyrosine-sensitive 3-deoxy-D-arabinoheptulosonate-7-phosphate (DAHP) and tyrosyl-tRNA synthetases on agarose carrying carboxyl-linked tyrosine.

@article{Gallopo1974PurificationOT,
  title={Purification of tyrosine-sensitive 3-deoxy-D-arabinoheptulosonate-7-phosphate (DAHP) and tyrosyl-tRNA synthetases on agarose carrying carboxyl-linked tyrosine.},
  author={A. Gallopo and P. Kotsiopoulos and S. Mohr},
  journal={Advances in experimental medicine and biology},
  year={1974},
  volume={42 0},
  pages={
          157-63
        }
}
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… Expand
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Affinity chromatography of a regulatory enzyme based on specific interaction with the effector.
TLDR
The tyrosine-sensitive 3-deoxy-D- arabino -heptulosonate-7-phosphate (DAHP) synthetase was retarded by the column relative to the bulk of the protein and was purified about 100-fold, while the phenylalanine- sensitive enzyme was not retarded. Expand
Purification of methionyl-tRNA synthetase from Escherichia coli K12 by affinity chromatography.
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High purified enzyme could be obtained in high yield by lowering the methionine substitution of the polymer to 4.5 μmol/ml gel and by introducing the affinity chromatography after the first DEAE-Sephadex step of the conventional purification procedure. Expand
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It is demonstrated that successful application of affinity chromatography in many cases will critically depend on placing the ligand at a considerable distance from the matrix backbone. Expand
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Partially digested s-RNA's were eluted from a column of methylated albumin of different salt concentration than untreated s- RNA's suggesting that relatively small alterations could be made in resistant s-RNAs without complete loss of acceptor activity for the two amino acids, phenylalanine and leucine. Expand