Monomeric C-mannosyl tryptophan is a degradation product of autophagy in cultured cells

Shiho Minakata, Yoko Inai, Shino Manabe, Kazuchika Nishitsuji, Yukishige Ito, Yoshito Ihara

Research output: Contribution to journalArticlepeer-review

Abstract

C-Mannosyl tryptophan (C-Man-Trp) is a unique glycosylated amino acid present in various eukaryotes. The C-Man-Trp structure can be found as a monomeric form or a part of post-translational modifications within polypeptide chains in living organisms. However, the mechanism of how monomeric C-Man-Trp is produced has not been fully investigated. In this study, we assessed levels of cellular C-Man-Trp by ultra performance liquid chromatography with a mass spectrometry assay system, and investigated whether the cellular C-Man-Trp is affected by autophagy induction. The intracellular C-Man-Trp level was significantly increased under serum and/or amino acid starvation in A549, HaCaT, HepG2, NIH3T3, and NRK49F cells. The increase in C-Man-Trp was also observed in NIH3T3 cells treated with rapamycin, an autophagy inducer. The up-regulation of C-Man-Trp caused by starvation was reversed by the inhibition of lysosomal enzymes. We further showed that C-Man-Trp is produced by incubating a synthetic C-mannosylated peptide (C-Man-Trp-Ser-Pro-Trp) or thrombospondin (TSP) in a lysosomal fraction that was prepared from a mouse liver, which provides supporting evidence that C-Man-Trp is a degradation product of the C-mannosylated peptide or protein following lysosome-related proteolysis. Taken together, we propose that the autophagic pathway is a novel pathway that at least partly contributes to intracellular C-Man-Trp production under certain conditions, such as nutrient starvation.

Original languageEnglish
Pages (from-to)635-645
Number of pages11
JournalGlycoconjugate Journal
Volume37
Issue number5
DOIs
Publication statusPublished - 2020 Oct 1

Keywords

  • Autophagy
  • C-mannosyl tryptophan
  • C-mannosylation
  • Glycosylation

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Cell Biology

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