The fibrinogen C-terminal domain is seldom C-mannosylated but its C-mannosylation is important for the secretion of microfibril-associated glycoprotein 4
Autor: | Takehiro Suzuki, Siro Simizu, Naoshi Dohmae, Kento Mori, Yoshiyuki Osada, Kazuki Miura, Hayato Mizuta |
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Rok vydání: | 2019 |
Předmět: |
0301 basic medicine
Biophysics Fibrinogen Biochemistry 03 medical and health sciences 0302 clinical medicine Protein Domains Cell Line Tumor medicine Humans Secretion Cell adhesion Molecular Biology Tissue homeostasis Glycoproteins chemistry.chemical_classification Extracellular Matrix Proteins Chemistry Cell biology carbohydrates (lipids) Protein Transport 030104 developmental biology Cell culture 030220 oncology & carcinogenesis Mannosylation HT1080 Glycoprotein Carrier Proteins Mannose medicine.drug |
Zdroj: | Biochimica et biophysica acta. General subjects. 1864(9) |
ISSN: | 1872-8006 |
Popis: | 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. |
Databáze: | OpenAIRE |
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