ATPase activity of non-ribosomal peptide synthetases.
Adenylation domains of non-ribosomal peptide synthetases (NRPS) catalyse the formation of aminoacyl adenylates, and in addition synthesize mono- and dinucleoside polyphosphates. Here, we show that NRPS systems furthermore contain an ATPase activity in the range of up to 2 P(i)/min. The hydrolysis rate by apo-tyrocidine synthetase 1 (apo-TY1) is enhanced in the presence of non-cognate amino acid substrates, correlating well with their structural features and the diminishing adenylation efficiency. A comparative analysis of the functional relevance of an analogous sequence motif in P-type ATPases and adenylate kinases (AK) allowed a putative assignment of the invariant aspartate residue from the TGDLA(V)R(K) core sequence in NRPS as the Mg(2+) binding site. Less pronounced variations in ATPase activity are observed in domains with relaxed amino acid specificity of gramicidin S synthetase 2 (GS2) and delta-(L-aminoadipyl)-L-cysteinyl-D-valine synthetase (ACVS), known to produce a set of substitutional variants of the respective peptide product. These results disclose new perspectives about the mode of substrate selection by NRPS.
Pavela-Vrancic
Maja
M
Department of Chemistry, Faculty of Natural Sciences, Mathematics and Education, University of Split, N Tesle 12, 21000 Split, Croatia.
Dieckmann
Ralf
R
von Döhren
Hans
H
D000251:Adenosine Triphosphatases / Q000737:chemistry* / Q000378:metabolism*; D000255:Adenosine Triphosphate / Q000737:chemistry / Q000378:metabolism; D000591:Amino Acid Isomerases / Q000737:chemistry; D004926:Escherichia coli / Q000378:metabolism; D006863:Hydrogen-Ion Concentration; D009097:Multienzyme Complexes / Q000737:chemistry; D009154:Mutation; D010453:Peptide Synthases / Q000096:biosynthesis* / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D010710:Phosphates / Q000032:analysis; D013053:Spectrophotometry; D013379:Substrate Specificity
D009097:Multienzyme Complexes; D010710:Phosphates; D000255:Adenosine Triphosphate; D000251:Adenosine Triphosphatases; D000591:Amino Acid Isomerases; D010453:Peptide Synthases; C020831:gramicidin-S-synthetase 2; C104385:non-ribosomal peptide synthase; C035180:tyrocidine synthetase