scholarly journals Active site-directed proteomic probes for adenylation domains in nonribosomal peptide synthetases

2015 ◽  
Vol 51 (12) ◽  
pp. 2262-2265 ◽  
Author(s):  
Sho Konno ◽  
Fumihiro Ishikawa ◽  
Takehiro Suzuki ◽  
Naoshi Dohmae ◽  
Michael D. Burkart ◽  
...  
Keyword(s):  
Natural Product ◽  
Active Site ◽  
Nonribosomal Peptide Synthetase ◽  
Protein Labeling ◽  
Nonribosomal Peptide ◽  
Nonribosomal Peptide Synthetases ◽  
Peptide Synthetases ◽  
Peptide Synthetase ◽  
A Domains

Active site-directed proteomic probes coupled to the 5′-O-N-(aminoacyl)sulfamoyladenosine (AMS) scaffold with a clickable benzophenone functionality selectively target nonribosomal peptide synthetase (NRPS) adenylation (A) domains in natural product producer proteomes by ligand-directed protein labeling.

scholarly journals Biosynthesis of Novel Pyoverdines by Domain Substitution in a Nonribosomal Peptide Synthetase of Pseudomonas aeruginosa

2014 ◽  
Vol 80 (18) ◽  
pp. 5723-5731 ◽  
Author(s):  
Mark J. Calcott ◽  
Jeremy G. Owen ◽  
Iain L. Lamont ◽  
David F. Ackerley
Keyword(s):  
Pseudomonas Aeruginosa ◽  
Peptide Bond ◽  
Nonribosomal Peptide Synthetase ◽  
Nonribosomal Peptide ◽  
Content Type ◽  
Threonine Residue ◽  
Peptide Synthetase ◽  
A Domains ◽  
High Yields ◽  
Acid Substrate

ABSTRACTPyoverdine is a fluorescent nonribosomal peptide siderophore made by fluorescent pseudomonads. ThePseudomonas aeruginosanonribosomal peptide synthetase (NRPS) PvdD contains two modules that each incorporate anl-threonine residue at the C-terminal end of pyoverdine. In an attempt to generate modified pyoverdine peptides, we substituted alternative-substrate-specifying adenylation (A) and peptide bond-catalyzing condensation (C) domains into the second module of PvdD. When just the A domain was substituted, the resulting strains produced only wild-type pyoverdine—at high levels if the introduced A domain specified threonine or at trace levels otherwise. The high levels of pyoverdine synthesis observed whenever the introduced A domain specified threonine indicated that these nonnative A domains were able to communicate effectively with the PvdD C domain. Moreover, the unexpected observation that non-threonine-specifying A domains nevertheless incorporated threonine into pyoverdine suggests that the native PvdD C domain exhibited stronger selectivity than these A domains for the incorporated amino acid substrate (i.e., misactivation of a threonine residue by the introduced A domains was more frequent than misincorporation of a nonthreonine residue by the PvdD C domain). In contrast, substitution of both the C and A domains of PvdD generated high yields of rationally modified pyoverdines in two instances, these pyoverdines having either a lysine or a serine residue in place of the terminal threonine. However, C-A domain substitution more commonly yielded a truncated peptide product, likely due to stalling of synthesis on a nonfunctional recombinant NRPS template.

A Multiple-Labeling Strategy for Nonribosomal Peptide Synthetases Using Active-Site-Directed Proteomic Probes for Adenylation Domains

ChemBioChem ◽  
2015 ◽  
Vol 16 (18) ◽  
pp. 2590-2594 ◽  
Author(s):  
Fumihiro Ishikawa ◽  
Takehiro Suzuki ◽  
Naoshi Dohmae ◽  
Hideaki Kakeya
Keyword(s):  
Active Site ◽  
Nonribosomal Peptide ◽  
Nonribosomal Peptide Synthetases ◽  
Peptide Synthetases

Probing the limits of interrupted adenylation domains by engineering a trifunctional enzyme capable of adenylation, N-, and S-methylation

2019 ◽  
Vol 17 (5) ◽  
pp. 1169-1175 ◽  
Author(s):  
Taylor A. Lundy ◽  
Shogo Mori ◽  
Sylvie Garneau-Tsodikova
Keyword(s):  
Nonribosomal Peptide ◽  
Nonribosomal Peptide Synthetases ◽  
Peptide Synthetases ◽  
A Domains

The adenylation (A) domains found in nonribosomal peptide synthetases (NRPSs) exhibit tremendous plasticity.

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