The fibrinogen C-terminal domain is seldom C-mannosylated but its C-mannosylation is important for the secretion of microfibril-associated glycoprotein 4

Yoshiyuki Osada, Takehiro Suzuki, Hayato Mizuta, Kento Mori, Kazuki Miura, Naoshi Dohmae, Siro Simizu

Research output: Contribution to journalArticlepeer-review

Abstract

Background: C-mannosylation is the one of glycosylations. Microfibril-associated glycoprotein 4 (MFAP4), an important protein for tissue homeostasis and cell adhesion, contains a consensus sequence of C-mannosylation in its fibrinogen C-terminal domain. In this study, we sought to demonstrate that fibrinogen C-terminal domain is a new substrate domain for C-mannosylation. Methods: We established an MFAP4-overexpresssing HT1080 cell line and purified recombinant MFAP4 protein from the conditioned medium for LC-MS/MS analysis. Subcellular localization of MFAP4 was observed under confocal fluorescence microscope. Results: We found that MFAP4 is C-mannosylated at Trp235 in the fibrinogen C-terminal domain by LC-MS/MS. To determine the functions of the C-mannosylation of MFAP4, we established a C-mannosylation-defective mutant MFAP4-overexpresssing HT1080 cell line and measured its secretion of MFAP4. The secretion of MFAP4 decreased significantly in the C-mannosylation-defective mutant MFAP4-overexpresssing cell line versus wild-type cells. Moreover, co-transfection experiments indicated that C-mannosylated MFAP4 accelerated its secretion. Conclusions: Our results demonstrate that the fibrinogen C-terminal domain is a novel C-mannosylation domain and that the C-mannosylation of MFAP4 is important for its secretion. General significance: These results suggest that C-mannosylation has a role for dominant effect for MFAP4 secretion.

Original languageEnglish
Article number129637
JournalBiochimica et Biophysica Acta - General Subjects
Volume1864
Issue number9
DOIs
Publication statusPublished - 2020 Sep

Keywords

  • C-mannosylation
  • Fibrinogen C-terminal domain
  • Glycosylation
  • Microfibril-associated glycoprotein 4

ASJC Scopus subject areas

  • Biophysics
  • Biochemistry
  • Molecular Biology

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