Preparation and Application of a Chemical Probe for Identifying the Targets of the Marine Cyclic Peptide Kapakahine A

Molecules. 2022 Feb 5;27(3):1072. doi: 10.3390/molecules27031072.

Abstract

Marine organisms are a rich source of bioactive secondary metabolites. Although many marine natural products with bioactivities have been isolated, successful elucidation of their mechanisms of action remains limited. In this study, we prepared a probe molecule based on the marine cyclic peptide kapakahine A (1) by introducing a linker with an azide terminal group, which enables the introduction of fluorescent groups for the effective monitoring of subcellular localization, or coupling to affinity beads for the pull-down of target proteins. The results of LC/MS/MS measurements, ProteinPilot analysis, and Western blotting suggest that kapakahine A interacts with the mitochondrial inner membrane proteins PHB1, PHB2, and ANT2, which is consistent with the results of the subcellular localization analysis using a fluorescent probe.

Keywords: chemical biology; chemical fluorescent probe; cyclic peptide; marine natural product; target protein.

MeSH terms

  • Adenine Nucleotide Translocator 2 / metabolism*
  • Animals
  • Aquatic Organisms / chemistry*
  • Cell Line
  • Chromatography, Liquid
  • Fluorescent Dyes / chemistry*
  • Mice
  • Molecular Structure
  • Peptides, Cyclic / chemistry
  • Peptides, Cyclic / pharmacology*
  • Prohibitins / metabolism*
  • Secondary Metabolism
  • Tandem Mass Spectrometry

Substances

  • Adenine Nucleotide Translocator 2
  • Fluorescent Dyes
  • Peptides, Cyclic
  • Prohibitins
  • kapakahine A