6.3.5.7 ATP + Asp-tRNAAsn + L-glutamine - Acidithiobacillus ferrooxidans ADP + phosphate + Asn-tRNAAsn + L-glutamate - ? 323522 6.3.5.7 ATP + Asp-tRNAAsn + L-glutamine - Helicobacter pylori ADP + phosphate + Asn-tRNAAsn + L-glutamate - ? 323522 6.3.5.7 ATP + Asp-tRNAAsn + L-glutamine identity elements used by GatCAB to discriminate tRNAAsn from tRNAAsp. GatCAB specifically binds Asp-tRNAAsn. Therefore, modified nucleotides do not play an essential role in GatCAB discrimination of Asp-tRNAAsn from Asp-tRNAAsp Methanothermobacter thermautotrophicus ADP + phosphate + Asn-tRNAAsn + L-glutamate - ? 323522 6.3.5.7 ATP + Asp-tRNAAsn + L-glutamine the enzyme transamidates Asp-tRNAAsn and Glu-tRNAGln with similar efficiency Helicobacter pylori ADP + phosphate + Asn-tRNAAsn + L-glutamate - ? 323522 6.3.5.7 ATP + Glu-tRNAGln + Asn - Chlamydomonas reinhardtii ADP + phosphate + Gln-tRNAGln + Asp - ? 323525 6.3.5.7 ATP + Glu-tRNAGln + Asn Asn is much less effective as amide donor than glutamine Bacillus subtilis ADP + phosphate + Gln-tRNAGln + Asp - ? 323525 6.3.5.7 ATP + Glu-tRNAGln + L-asparagine - Bacillus subtilis ADP + phosphate + Gln-tRNAGln + L-glutamate - ? 323523 6.3.5.7 ATP + Glu-tRNAGln + L-glutamine - Acidithiobacillus ferrooxidans ADP + phosphate + Gln-tRNAGln + L-glutamate - ? 323521 6.3.5.7 ATP + Glu-tRNAGln + L-glutamine - Chlamydomonas reinhardtii ADP + phosphate + Gln-tRNAGln + L-glutamate - ? 323521 6.3.5.7 ATP + Glu-tRNAGln + L-glutamine - Bacillus subtilis ADP + phosphate + Gln-tRNAGln + L-glutamate - ? 323521 6.3.5.7 ATP + Glu-tRNAGln + L-glutamine - Geobacillus stearothermophilus ADP + phosphate + Gln-tRNAGln + L-glutamate - ? 323521 6.3.5.7 ATP + Glu-tRNAGln + L-glutamine - Methanothermobacter thermautotrophicus ADP + phosphate + Gln-tRNAGln + L-glutamate - ? 323521 6.3.5.7 ATP + Glu-tRNAGln + L-glutamine - Helicobacter pylori ADP + phosphate + Gln-tRNAGln + L-glutamate - ? 323521 6.3.5.7 ATP + Glu-tRNAGln + L-glutamine - Homo sapiens ADP + phosphate + Gln-tRNAGln + L-glutamate - ? 323521 6.3.5.7 ATP + Glu-tRNAGln + L-glutamine Ser176A is the active-site nucleophile for facilitating Gln hydrolysis by the enzyme Streptococcus pyogenes ADP + phosphate + Gln-tRNAGln + L-glutamate - ? 323521 6.3.5.7 ATP + Glu-tRNAGln + L-glutamine the amidation of Glu-tRNAGln proceeds via a gamma-phosphorylated intermediate Bacillus subtilis ADP + phosphate + Gln-tRNAGln + L-glutamate - ? 323521 6.3.5.7 ATP + Glu-tRNAGln + L-glutamine organisms lacking Gln-tRNA synthetase produce Gln-tRNAGln from misacylated Glu-tRNAGln through the transamidation activity of Glu-tRNAGln amidotransferase. The enzyme hydrolyzes Gln to Glu and NH3, using the latter product to transamidate Glu-tRNAGln in concert with ATP hydrolysis Streptococcus pyogenes ADP + phosphate + Gln-tRNAGln + L-glutamate - ? 323521 6.3.5.7 ATP + Glu-tRNAGln + L-glutamine the enzyme produces Gln-tRNAGln required for plastidal protein biosynthesis Tetradesmus obliquus ADP + phosphate + Gln-tRNAGln + L-glutamate - ? 323521 6.3.5.7 ATP + Glu-tRNAGln + L-glutamine disruption of this operon is lethal. Transamidation is the only pathway to Gln-tRNAGln in Bacillus subtilis. The enzyme furnishes a means for formation of correctly charged Gln-tRNAGln through the transamidation of misacylated Glu-tRNAGln, functionally replacing the lack of glutaminyl-tRNA synthetase activity in Gram-positive eubacteria, cyanobacteria, archaea and organelles Bacillus subtilis ADP + phosphate + Gln-tRNAGln + L-glutamate - ? 323521 6.3.5.7 ATP + Glu-tRNAGln + L-glutamine GatDE is a heterodimeric amidotransferase. GatD acts as a glutaminase but only in the presence of both Glu-tRNAGln and the other subunit, GatE. The fact that only Glu-tRNAGln but not tRNA Gln could activate the glutaminase activity of GatD suggests that glutamine hydrolysis is coupled tightly to transamidation. GatE is a Glu-tRNAGln kinase that activates Glu-tRNAGln via gamma-phosphorylation Methanothermobacter thermautotrophicus ADP + phosphate + Gln-tRNAGln + L-glutamate - ? 323521 6.3.5.7 ATP + Glu-tRNAGln + L-glutamine the enzyme transamidates Asp-tRNAAsn and Glu-tRNAGln with similar efficiency. GatCAB uses the amide donor glutamine 129fold more efficiently than asparagine Helicobacter pylori ADP + phosphate + Gln-tRNAGln + L-glutamate - ? 323521 6.3.5.7 ATP + Glu-tRNAGln + L-glutamine + H2O - Helicobacter pylori ADP + phosphate + Gln-tRNAGln + L-glutamate - ? 427380 6.3.5.7 ATP + Glu-tRNAGln + L-glutamine + H2O - Thermotoga maritima ADP + phosphate + Gln-tRNAGln + L-glutamate - ? 427380 6.3.5.7 ATP + Glu-tRNAGln + NH4Cl - Chlamydomonas reinhardtii ADP + phosphate + Gln-tRNAGln + ? - ? 323526 6.3.5.7 ATP + Glu-tRNAGln + NH4Cl NH4Cl is much less effective as amide donor than glutamine Bacillus subtilis ADP + phosphate + Gln-tRNAGln + ? - ? 323526 6.3.5.7 ATP + L-aspartyl-tRNAAsn + L-glutamine + H2O - Helicobacter pylori ADP + phosphate + L-asparaginyl-tRNAAsn + L-glutamate - ? 453780 6.3.5.7 ATP + L-glutamate + tRNAGln1(UUG) the enzyme is active toward the two tRNAGln isoacceptors, but with a significant catalytic preference for tRNAGln2(CUG). The less active tRNAGln1(UUG) responds to the less common CAA codon for Gln Methanothermobacter thermautotrophicus AMP + diphosphate + L-glutamyl-tRNAGln1(UUG) - ? 428282 6.3.5.7 ATP + L-glutamate + tRNAGln2(CUG) the enzyme is active toward the two tRNAGln isoacceptors, but with a significant catalytic preference for tRNAGln2(CUG). The less active tRNAGln1(UUG) responds to the less common CAA codon for Gln. The wild-type enzyme shows a 24fold catalytic preference for tRNAGln2 over tRNAGlu Methanothermobacter thermautotrophicus AMP + diphosphate + L-glutamyl-tRNAGln2(CUG) - ? 428283 6.3.5.7 ATP + L-glutamate + tRNAGlu - Methanothermobacter thermautotrophicus AMP + diphosphate + L-glutamyl-tRNAGlu - ? 73 6.3.5.7 ATP + L-glutamyl-tRNAGln + L-glutamine - Saccharomyces cerevisiae ADP + phosphate + L-glutaminyl-tRNAGln + L-glutamate - ? 443154 6.3.5.7 ATP + L-glutamyl-tRNAGln + L-glutamine - Plasmodium falciparum ADP + phosphate + L-glutaminyl-tRNAGln + L-glutamate - ? 443154 6.3.5.7 ATP + L-glutamyl-tRNAGln + L-glutamine - Plasmodium berghei ADP + phosphate + L-glutaminyl-tRNAGln + L-glutamate - ? 443154 6.3.5.7 ATP + L-glutamyl-tRNAGln + L-glutamine - Plasmodium berghei ANKA ADP + phosphate + L-glutaminyl-tRNAGln + L-glutamate - ? 443154 6.3.5.7 ATP + L-glutamyl-tRNAGln + L-glutamine - Saccharomyces cerevisiae ATCC 204508 / S288c ADP + phosphate + L-glutaminyl-tRNAGln + L-glutamate - ? 443154 6.3.5.7 ATP-gammaS + Glu-tRNAGln + L-glutamine - Streptococcus pyogenes ? + phosphate + Gln-tRNAGln + L-glutamate - ? 323524 6.3.5.7 additional information in absence of the amido acceptor, Glu-tRNAGln, the enzyme has basal glutaminase activity that is unaffected by ATP Streptococcus pyogenes ? - ? 89 6.3.5.7 additional information the enzyme possesses low glutaminase activity Chlamydomonas reinhardtii ? - ? 89 6.3.5.7 additional information the association of archaeal glutamyl-tRNA synthetase (ND-GluRS) with GatDE sequesters the tRNA synthetase for Gln-tRNAGln formation, with GatDE reducing the affinity of glutamyl-tRNA synthetase (ND-GluRS) for tRNAGlu 13fold Methanothermobacter thermautotrophicus ? - ? 89 6.3.5.7 additional information no substrate: L-glutamate Plasmodium falciparum ? - ? 89 6.3.5.7 additional information no substrate: L-glutamate Plasmodium berghei ? - ? 89 6.3.5.7 additional information no substrate: L-glutamate Plasmodium berghei ANKA ? - ? 89