title: Isolation and characterization of a guanine insertion enzyme, a specific tRNA transglycosylase, from Escherichia coli.
abstract: A guanine insertion enzyme (tRNA transglycosylase) was purified to a homogeneous state from Escherichia coli B by ammonium sulfate fractionation and DEAE-cellulose, DEAE-Sephadex A-50, phosphocellulose, and Sephadex G-200 column chromatographies. The molecular weight of the enzyme, which appeared to be a single polypeptide, was 4.6 X 10(4) by sodium dodecyl sulfate gel electrophoresis. The enzyme catalyzes exchange of guanine with guanine located in the first position of the anticodon of tRNATyr, tRNAHis, tRNAAsn, and tRNAAsp, but unlike the enzymes isolated from rabbit reticulocytes and Ehrlich ascites tumor cells it does not catalyze the exchange of guanine with queuine (7-(3,4-trans-4,5-cis-dihydroxy-1-cyclopenten-3-ylaminomethyl)-7-deazaguanine) present in these tRNAs. The pH optimum of the reaction was 7.0, and the pH1 value was 4.6 to 4.8. The reaction required Mg2+ ion. 7-Methylguanine inhibited guanine insertion, but the other purine analogues tested were not inhibitory and could not replace guanine.20
mesh_terms: D001483:Base Sequence; D004926:Escherichia coli / Q000201:enzymology*; D006147:Guanine; D007700:Kinetics; D009843:Oligoribonucleotides / Q000032:analysis; D010430:Pentosyltransferases; D012343:RNA, Transfer; D006163:Ribonuclease T1; D014166:Transferases / Q000302:isolation & purification / Q000378:metabolism*; D014443:Tyrosine
chemical_list: D009843:Oligoribonucleotides; D014443:Tyrosine; D006147:Guanine; D012343:RNA, Transfer; D014166:Transferases; D010430:Pentosyltransferases; C020837:queuine tRNA-ribosyltransferase; D006163:Ribonuclease T1