Synthesis and NMR Characterization of the Prenylated Peptide, a-Factor

Bader, Taysir K.; Rappe, Todd M.; Veglia, Gianlugi; Distefano, Mark D.

doi:10.1016/bs.mie.2018.09.025

Cited by 2 publications

(3 citation statements)

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“…a-Factor has been extensively studied for its three posttranslational modifications (isoprenylation, proteolysis, and carboxyl methylation) that are required for proper mating between two haploid yeast ( S. cerevisiae ) cells. − a-Factor precursors 9a and 10a containing VIA and Cys-COOH C-termini were synthesized by standard solid phase peptide synthetic methods. a-Factor precursor 11a with a C-terminal methyl ester was prepared using a side chain anchoring methodology where Fmoc-Cys-OMe linked to a trityl resin via its thiol group (Figure S7) was employed for subsequent solid phase peptide synthesis. , Subsequent peptides were then prenylated chemically with trans , trans -farnesyl bromide or the corresponding chloride precursor used to prepare AzoFPP-1 at pH 5.0 in the presence of Zn(OAc) 2 and NaI. These conditions were optimized (Figure S8) based on previously reported procedures. − Peptides containing a VIA ( 9b and 9c ), Cys-COOH ( 10b and 10c ), or Cys-COMe ( 11b and 11c ) termini were obtained in this manner.…”

Section: Resultsmentioning

confidence: 99%

“…a-Factor precursor 11a with a C-terminal methyl ester was prepared using a side chain anchoring methodology where Fmoc-Cys-OMe linked to a trityl resin via its thiol group (Figure S7) was employed for subsequent solid phase peptide synthesis. , Subsequent peptides were then prenylated chemically with trans , trans -farnesyl bromide or the corresponding chloride precursor used to prepare AzoFPP-1 at pH 5.0 in the presence of Zn(OAc) 2 and NaI. These conditions were optimized (Figure S8) based on previously reported procedures. − Peptides containing a VIA ( 9b and 9c ), Cys-COOH ( 10b and 10c ), or Cys-COMe ( 11b and 11c ) termini were obtained in this manner. Using these model peptides, each processing enzyme was assayed for activity with its respective a-factor substrate in either the trans -form (dark) or cis -form (after UV-A irradiation) for light-dependent conversion.…”

Section: Resultsmentioning

confidence: 99%

Section: T H Imentioning

confidence: 99%

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Photoswitchable Isoprenoid Lipids Enable Optical Control of Peptide Lipidation

et al. 2022

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Photoswitchable lipids have emerged as attractive tools for the optical control of lipid bioactivity, metabolism, and biophysical properties. Their design is typically based on the incorporation of an azobenzene photoswitch into the hydrophobic lipid tail, which can be switched between its trans- and cis-form using two different wavelengths of light. While glycero- and sphingolipids have been successfully designed to be photoswitchable, isoprenoid lipids have not yet been investigated. Herein, we describe the development of photoswitchable analogs of an isoprenoid lipid and systematically assess their potential for the optical control of various steps in the isoprenylation processing pathway of CaaX proteins in Saccharomyces cerevisiae. One photoswitchable analog of farnesyl diphosphate (AzoFPP-1) allowed effective optical control of substrate prenylation by farnesyltransferase. The subsequent steps of isoprenylation processing (proteolysis by either Ste24 or Rce1 and carboxyl methylation by Ste14) were less affected by photoisomerization of the group introduced into the lipid moiety of the substrate a-factor, a mating pheromone from yeast. We assessed both proteolysis and methylation of the a-factor analogs in vitro and the bioactivity of a fully processed a-factor analog containing the photoswitch, exogenously added to cognate yeast cells. Combined, these data describe the first successful conversion of an isoprenoid lipid into a photolipid and suggest the utility of this approach for the optical control of protein prenylation.

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