An Intrabody against B-Cell Receptor-Associated Protein 31 (BAP31) Suppresses the Glycosylation of the Epithelial Cell-Adhesion Molecule (EpCAM) via Affecting the Formation of the Sec61-Translocon-Associated Protein (TRAP) Complex.

Abstract

The epithelial cell-adhesion molecule (EpCAM) is hyperglycosylated in carcinoma tissue and the oncogenic function of EpCAM primarily depends on the degree of glycosylation. Inhibiting EpCAM glycosylation is expected to have an inhibitory effect on cancer. We analyzed the relationship of BAP31 with 84 kinds of tumor-associated antigens and found that BAP31 is positively correlated with the protein level of EpCAM. Triple mutations of EpCAM N76/111/198A, which are no longer modified by glycosylation, were constructed to determine whether BAP31 has an effect on the glycosylation of EpCAM. Plasmids containing different C-termini of BAP31 were constructed to identify the regions of BAP31 that affects EpCAM glycosylation. Antibodies against BAP31 (165-205) were screened from a human phage single-domain antibody library and the effect of the antibody (VH-F12) on EpCAM glycosylation and anticancer was investigated. BAP31 increases protein levels of EpCAM by promoting its glycosylation. The amino acid region from 165 to 205 in BAP31 plays an important role in regulating the glycosylation of EpCAM. The antibody VH-F12 significantly inhibited glycosylation of EpCAM which, subsequently, reduced the adhesion of gastric cancer cells, inducing cytotoxic autophagy, inhibiting the AKT-PI3K-mTOR signaling pathway, and, finally, resulting in proliferation inhibition both in vitro and in vivo. Finally, we clarified that BAP31 plays a key role in promoting N-glycosylation of EpCAM by affecting the Sec61 translocation channels. Altogether, these data implied that BAP31 regulates the N-glycosylation of EpCAM and may represent a potential therapeutic target for cancer therapy.

• Authors: Tianyi Wang, Changli Wang, Jiyu Wang, Bing Wang
• Journal: International journal of molecular sciences (Int J Mol Sci) Vol. 24 Issue 19 (Sep 30 2023) ISSN: 1422-0067 [Electronic] Switzerland
• PMID: 37834237 (Publication Type: Journal Article)

Chemical References

• Antigens, Neoplasm
• Epithelial Cell Adhesion Molecule
• Receptors, Antigen, B-Cell
• SEC Translocation Channels
• signal sequence receptor
• Antibodies
• BCAP31 protein, human
• Membrane Proteins

Topics

• Humans
• Antigens, Neoplasm (immunology)
• Carcinoma
• Cell Line, Tumor
• Epithelial Cell Adhesion Molecule (genetics, metabolism)
• Epithelial Cells (metabolism)
• Glycosylation
• Receptors, Antigen, B-Cell (metabolism)
• SEC Translocation Channels (metabolism)
• Antibodies
• Membrane Proteins (immunology)