Analytical strategy for determination of active site sequences in aminoacyl-tRNA synthetases.
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.
Beauvallet
C
C
Laboratoire de Biochimie, Ecole Polytechnique, Palaiseau, France.
Hountondji
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C
Schmitter
J M
JM
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
D010455:Peptides; D012822:Silicon Dioxide; C049272:Spherogel TSK 2000SW; D012260:Ribonucleases; D002918:Chymotrypsin; D000604:Amino Acyl-tRNA Synthetases