A Single Amino Acid Switch Alters the Isoprene Donor Specificity in Ribosomally Synthesized and Post-Translationally Modified Peptide Prenyltransferases

Journal Article · 20 June 2018 · Journal of the American Chemical Society

DOI: https://doi.org/10.1021/jacs.8b05187 · OSTI ID:1544874

Estrada, Paola ^[1]; Morita, Maho ^[2]; Hao, Yue ^[3]; Schmidt, Eric W.  ^[2]; Nair, Satish K.  ^[1]

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1. Univ. of Illinois at Urbana-Champaign, IL (United States)
2. Univ. of Utah, Salt Lake City, UT (United States)
3. Univ. of Illinois at Urbana-Champaign, IL (United States); Amgen Pharmaceuticals, Cambridge, MA (United States)

Mutation at a single amino acid alters the isoprene donor specificity of prenyltransferases involved in the modification of ribosomally synthesized and post-translationally modified peptides (RiPPs). Though most characterized RiPP prenyltransferases carry out the regiospecific transfer of C₅ dimethylallyl donor to the side chain atoms on macrocyclic acceptor substrates, the elucidation of the cyanobactin natural product piricyclamide 70005E1 identifies an O-geranyl modification on Tyr, a reaction with little prior biochemical precedence. Reconstitution and kinetic studies of the presumptive geranyltransferase PirF shows that the enzyme utilizes a C₁₀ donor, with no C₅ transferase activity. The crystal structure of PirF reveals a single amino acid difference in the vicinity of the isoprene-binding pocket, relative to the C₅ utilizing enzymes. Remarkably, only a single amino acid mutation is necessary to completely switch the donor specificity from a C₅ to a C₁₀ prenyltransferase, and vice versa. Lastly, we demonstrate that these enzymes may be used for the chemospecific attachment of C₅ or C₁₀ lipid groups on lanthipeptides, an unrelated class of RiPP natural products. Overall, these studies represent a rare example where prenyl donor specificity can be discretely altered, which expands the arsenal of synthetic biology tools for tuning biological activities of peptide natural products.

• Research Organization: Advanced Photon Source (APS), Argonne National Laboratory (ANL), Argonne, IL (US)
• Sponsoring Organization: National Institutes of Health (NIH)
• OSTI ID: 1544874
• Journal Information: Journal of the American Chemical Society, Journal Name: Journal of the American Chemical Society Journal Issue: 26 Vol. 140; ISSN 0002-7863
• Publisher: American Chemical Society (ACS)Copyright Statement
• Country of Publication: United States
• Language: ENGLISH

Related Subjects

37 INORGANIC, ORGANIC, PHYSICAL, AND ANALYTICAL CHEMISTRY
Peptides and proteins
chemical specificity
chemical structure
crystal structure
monomers