title: Analytical strategy for determination of active site sequences in aminoacyl-tRNA synthetases.
abstract: Affinity labelling with radioactive, periodate-oxidized tRNA has been used to investigate the structures of tRNA-binding sites in Escherichia coli aminoacyl-tRNA synthetases. Labelled peptides were isolated by means of a combination of techniques involving chymotryptic digestion of the enzyme, gel filtration, ribonuclease digestion of tRNA, chromatography on a TSK 2000 column and reversed-phase chromatography. An isocratic phenylthiohydantoin identification system has been interfaced to a sequencer, allowing the characterization of modified lysine residues by means of both chromatographic retention and liquid scintillation counting.
authors.affiliation: Laboratoire de Biochimie, Ecole Polytechnique, Palaiseau, France.
authors.forename: C
authors.initials: C
authors.lastname: Beauvallet
authors.forename: C
authors.initials: C
authors.lastname: Hountondji
authors.forename: J M
authors.initials: JM
authors.lastname: Schmitter
mesh_terms: D000604:Amino Acyl-tRNA Synthetases / Q000032:analysis*; D001665:Binding Sites; D002846:Chromatography, Affinity; D002850:Chromatography, Gel; D002853:Chromatography, Liquid; D002918:Chymotrypsin; D004926:Escherichia coli / Q000201:enzymology; D006868:Hydrolysis; D010455:Peptides / Q000032:analysis; D012260:Ribonucleases / Q000378:metabolism; D012822:Silicon Dioxide; D013056:Spectrophotometry, Ultraviolet
chemical_list: D010455:Peptides; D012822:Silicon Dioxide; C049272:Spherogel TSK 2000SW; D012260:Ribonucleases; D002918:Chymotrypsin; D000604:Amino Acyl-tRNA Synthetases