title: Unusual resistance of peptidyl transferase to protein extraction procedures.
abstract: Peptidyl transferase, the ribosomal activity responsible for catalysis of peptide bond formation, is resistant to vigorous procedures that are conventionally employed to remove proteins from protein-nucleic acid complexes. When the "fragment reaction" was used as a model assay for peptide bond formation, Escherichia coli ribosomes or 50S subunits retained 20 to 40 percent activity after extensive treatment with proteinase K and SDS, but lost activity after extraction with phenol or exposure to EDTA. Ribosomes from the thermophilic eubacterium Thermus aquaticus remained more than 80 percent active after treatment with proteinase K and SDS, which was followed by vigorous extraction with phenol. This activity is attributable to peptidyl transferase, as judged by specific inhibition by the peptidyl transferase-specific antibiotics chloramphenicol and carbomycin. In contrast, activity is abolished by treatment with ribonuclease T1. These findings support the possibility that 23S ribosomal RNA participates in the peptidyl transferase function.
authors.affiliation: Sinsheimer Laboratories, University of California, Santa Cruz.
authors.forename: H F
authors.initials: HF
authors.lastname: Noller
authors.forename: V
authors.initials: V
authors.lastname: Hoffarth
authors.forename: L
authors.initials: L
authors.lastname: Zimniak
mesh_terms: D004591:Electrophoresis, Polyacrylamide Gel; D004926:Escherichia coli / Q000201:enzymology*; D046911:Macromolecular Substances; D008970:Molecular Weight; D010458:Peptidyl Transferases / Q000302:isolation & purification* / Q000378:metabolism*; D011691:Puromycin / Q000378:metabolism; D012338:RNA, Ribosomal, 23S / Q000302:isolation & purification* / Q000378:metabolism*; D012270:Ribosomes / Q000201:enzymology*; D013462:Sulfur Radioisotopes; D013824:Thermus / Q000201:enzymology*
chemical_list: D046911:Macromolecular Substances; D012338:RNA, Ribosomal, 23S; D013462:Sulfur Radioisotopes; D011691:Puromycin; D010458:Peptidyl Transferases