Preoperative isolation of a reversible protein-protein crosslink generated in 50 S subunits of Escherichia coli ribosomes and identification of its components.
The reversible crosslinked protein complex L13-L21 was isolated from Escherichia coli ribosomes in milligram amounts. Tight couples (70 S) were crosslinked with the bifunctional reagent diepoxybutane. The reacted ribosomes were separated into their subunits by sucrose gradient centrifugation in the presence of 1 mM magnesium. The crosslinked subunits were analyzed by symmetrical two-dimensional gel electrophoresis. One crosslinked protein complex detected within the large subunit was purified by salt extraction, acidic acid extraction, and ion-exchange chromatography. Two-dimensional gel electrophoresis and immunological results established L13 and L21 as the components of this crosslink.
Schönfeld
H J
HJ
Max-Planck-Institut für Molekulare Genetik, Abteilung Wittmann, Berlin, Federal Republic of Germany.
Foulaki
K
K
D001426:Bacterial Proteins / Q000302:isolation & purification*; D002852:Chromatography, Ion Exchange; D003432:Cross-Linking Reagents; D004591:Electrophoresis, Polyacrylamide Gel; D004852:Epoxy Compounds; D004926:Escherichia coli / Q000032:analysis*; D005455:Fluorescent Antibody Technique; D007118:Immunoassay; D012269:Ribosomal Proteins / Q000302:isolation & purification*; D012270:Ribosomes / Q000032:analysis*
D001426:Bacterial Proteins; D003432:Cross-Linking Reagents; D004852:Epoxy Compounds; D012269:Ribosomal Proteins; C007366:diepoxybutane