Evidence for a monomeric structure of nonribosomal Peptide synthetases.
Nonribosomal peptide synthetases (NRPS) are multimodular biocatalysts that bacteria and fungi use to assemble many complex peptides with broad biological activities. The same modular enzymatic assembly line principles are found in fatty acid synthases (FAS), polyketide synthases (PKS), and most recently in hybrid NRPS/PKS multienzymes. FAS as well as PKS are known to function as homodimeric enzyme complexes, raising the question of whether NRPS may also act as homodimers. To test this hypothesis, biophysical methods (size exclusion chromatography, analytical equilibrium ultracentrifugation, and chemical crosslinking) and biochemical methods (two-affinity-tag-system and complementation studies with enzymes being inactivated in different catalytic domains) were applied to NRPS subunits from the gramicidin S (GrsA-ATE), tyrocidine (TycB(1)-CAT and TycB(2-3)-AT.CATE), and enterobactin (EntF-CATTe) biosynthetic systems. These methods had revealed the dimeric structure of FAS and PKS previously, but all three NRPS systems investigated are functionally active as monomers.
D000345:Affinity Labels / Q000737:chemistry / Q000378:metabolism; D020134:Catalytic Domain; D002623:Chemistry Techniques, Analytical / Q000379:methods; D003432:Cross-Linking Reagents; D004758:Enterobactin / Q000737:chemistry / Q000378:metabolism; D006096:Gramicidin / Q000737:chemistry / Q000378:metabolism; D007700:Kinetics; D008958:Models, Molecular; D016297:Mutagenesis, Site-Directed; D010453:Peptide Synthases / Q000737:chemistry* / Q000235:genetics; D011487:Protein Conformation; D021122:Protein Subunits; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism; D014440:Tyrocidine / Q000737:chemistry / Q000378:metabolism; D014461:Ultracentrifugation / Q000379:methods / Q000706:statistics & numerical data
D000345:Affinity Labels; D003432:Cross-Linking Reagents; D021122:Protein Subunits; D011994:Recombinant Proteins; D014440:Tyrocidine; D006096:Gramicidin; D004758:Enterobactin; D010453:Peptide Synthases; C104385:non-ribosomal peptide synthase