title: Insights into methyltransferase specificity and bioactivity of derivatives of the antibiotic plantazolicin.
abstract: Peptide antibiotics represent a class of conformationally constrained natural products of growing pharmaceutical interest. Plantazolicin (PZN) is a linear, polyheterocyclic natural product with highly selective and potent activity against the anthrax-causing bacterium, Bacillus anthracis. The bioactivity of PZN is contingent on dimethylation of its N-terminal Arg residue by an S-adenosylmethionine-dependent methyltransferase. Here, we explore the substrate tolerances of two homologous PZN methyltransferases by carrying out kinetic analyses of the enzymes against a synthetic panel of truncated PZN analogs containing the N-terminal Arg residue. X-ray cocrystal structures of the PZN methyltransferases with each of these heterocycle-containing substrates provide a rationale for understanding the strict substrate specificity of these enzymes. Kinetic studies of structure-guided, site-specific variants allowed for the assignment of residues governing catalysis and substrate scope. Microbiological testing further revealed that upon dimethylation of the N-terminal Arg, a pentaheterocyclized PZN analog retained potent anti-B. anthracis activity, nearly equal to that of full-length PZN. These studies may be useful in the biosynthetic engineering of natural product analogs with different bioactivity profiles, as demonstrated by our identification of a truncated plantazolicin derivative that is active against methicillin-resistant Staphylococcus aureus (MRSA).
authors: Department of Biochemistry, University of Illinois at Urbana-Champaign, Urbana, IL 61801.
authors: Yue
authors: Y
authors: Hao
authors: Department of Chemistry, University of Illinois at Urbana-Champaign, Urbana, IL 61801.
authors: Patricia M B
authors: PMB
authors: Saint-Vincent
authors: Department of Chemistry, University of Illinois at Urbana-Champaign, Urbana, IL 61801.
authors: Abhishek
authors: A
authors: Sharma
authors: Department of Chemistry, University of Illinois at Urbana-Champaign, Urbana, IL 61801.|Department of Microbiology, University of Illinois at Urbana-Champaign, Urbana, IL 61801.|Institute for Genomic Biology, University of Illinois at Urbana-Champaign, Urbana, IL 61801.
authors: Douglas A
authors: DA
authors: Mitchell
authors: Department of Biochemistry, University of Illinois at Urbana-Champaign, Urbana, IL 61801.|Department of Chemistry, University of Illinois at Urbana-Champaign, Urbana, IL 61801.|Institute for Genomic Biology, University of Illinois at Urbana-Champaign, Urbana, IL 61801.|Center for Biophysics and Computational Biology, University of Illinois at Urbana-Champaign, Urbana, IL 61801.
authors: Satish K
authors: SK
authors: Nair
mesh_terms: D000595:Amino Acid Sequence; D000900:Anti-Bacterial Agents / Q000737:chemistry / Q000378:metabolism / Q000494:pharmacology*; D018360:Crystallography, X-Ray; D007700:Kinetics; D008780:Methyltransferases / Q000737:chemistry / Q000378:metabolism*; D008969:Molecular Sequence Data; D015394:Molecular Structure; D009842:Oligopeptides / Q000378:metabolism / Q000494:pharmacology*; D017386:Sequence Homology, Amino Acid; D013379:Substrate Specificity
chemical_list: D000900:Anti-Bacterial Agents; D009842:Oligopeptides; C560834:plantazolicin A; D008780:Methyltransferases
references: Org Lett. 2011 Jun 17;13(12):2996-9
references: 21568297
references: Science. 1996 Nov 15;274(5290):1188-93
references: 8895467
references: Curr Opin Chem Biol. 2011 Jun;15(3):369-78
references: 21429787
references: Chem Commun (Camb). 2013 Nov 25;49(91):10703-5
references: 24100616
references: J Appl Crystallogr. 2007 Aug 1;40(Pt 4):658-674
references: 19461840
references: Acta Crystallogr D Biol Crystallogr. 2010 Jan;66(Pt 1):12-21
references: 20057044
references: J Bacteriol. 2011 May;193(10):2510-6
references: 21421760
references: Acta Crystallogr D Biol Crystallogr. 2014 Aug;70(Pt 8):2204-16
references: 25084339
references: FEBS Lett. 2012 Sep 21;586(19):3391-7
references: 22841713
references: Nat Prod Rep. 2012 Oct;29(10):1238-50
references: 22850796
references: J Mol Biol. 2008 Apr 18;378(1):154-64
references: 18342334
references: Acta Crystallogr A. 2003 May;59(Pt 3):228-34
references: 12714773
references: Nat Prod Rep. 2013 Jan;30(1):108-60
references: 23165928
references: Chembiochem. 2010 Jul 5;11(10):1413-21
references: 20540059
references: J Bacteriol. 2011 Jan;193(1):215-24
references: 20971906
references: Biochemistry. 1999 Apr 13;38(15):4768-81
references: 10200165
references: Proc Natl Acad Sci U S A. 2013 Aug 6;110(32):12954-9
references: 23878226
references: Proc Natl Acad Sci U S A. 2010 Oct 12;107(41):17557-62
references: 20876132
references: ACS Chem Biol. 2011 Dec 16;6(12):1307-13
references: 21950656
references: J Clin Pathol. 2003 Mar;56(3):182-7
references: 12610093
references: Methods. 2011 Sep;55(1):94-106
references: 21821126
references: Acta Crystallogr D Biol Crystallogr. 2011 Apr;67(Pt 4):355-67
references: 21460454
references: J Biomol NMR. 1996 Dec;8(4):477-86
references: 9008363
references: Chembiochem. 2006 May;7(5):795-804
references: 16575939
references: Acta Crystallogr D Biol Crystallogr. 2004 Dec;60(Pt 12 Pt 1):2126-32
references: 15572765
references: ACS Chem Biol. 2013 Sep 20;8(9):1998-2008
references: 23823732
references: Chem Biol. 2005 May;12(5):589-93
references: 15911379
references: Org Lett. 2013 Oct 4;15(19):5076-9
references: 24063702