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Article: Epitope Mapping of Antibodies Suggests the Novel Membrane Topology of B-Cell Receptor Associated Protein 31 on the Cell Surface of Embryonic Stem Cells: The Novel Membrane Topology of BAP31

TitleEpitope Mapping of Antibodies Suggests the Novel Membrane Topology of B-Cell Receptor Associated Protein 31 on the Cell Surface of Embryonic Stem Cells: The Novel Membrane Topology of BAP31
Authors
Issue Date2015
Citation
PLOS ONE, 2015, v. 10 n. 6, p. e0130670 How to Cite?
AbstractWhen located in the endoplasmic reticulum (ER) membrane, B-cell receptor associated protein 31 (BAP31) is involved in the export of secreted proteins from the ER to the plasma membrane. In a previous study, we generated two monoclonal antibodies (mAbs), 297-D4 and 144-A8, that bound to surface molecules on human embryonic stem cells (hESCs), but not to surface molecules on mouse embryonic stem cells (mESCs). Subsequent studies revealed that the mAbs recognized BAP31 on the surface of hESCs. To investigate the membrane topology of BAP31 on the cell surface, we first examined the epitope specificity of 297-D4 and 144-A8, as well as a polyclonal anti-BAP31 antibody (alpha-BAP31). We generated a series of GST-fused BAP31 mutant proteins in which BAP31 was serially deleted at the C-terminus. GST-fused BAP31 mutant proteins were then screened to identify the epitopes targeted by the antibodies. Both 297-D4 and 144-A8 recognized C-terminal residues 208-217, while alpha-BAP31 recognized C-terminal residues 165-246, of BAP31 on hESCs, suggesting that the C-terminal domain of BAP31 is exposed on the cell surface. The polyclonal antibody alpha-BAP31 bound to mESCs, which confirmed that the C-terminal domain of BAP31 is also exposed on the surface of these cells. Our results show for the first time the novel membrane topology of cell surface-expressed BAP31 as the extracellular exposure of the BAP31 C-terminal domain was not predicted from previous studies.
Persistent Identifierhttp://hdl.handle.net/10722/211677
ISSN
2015 Impact Factor: 3.057
2015 SCImago Journal Rankings: 1.395
ISI Accession Number ID

 

DC FieldValueLanguage
dc.contributor.authorKim, WT-
dc.contributor.authorChoi, HS-
dc.contributor.authorHwang, HJ-
dc.contributor.authorJung, HS-
dc.contributor.authorRyu, CJ-
dc.contributor.authorSchnell, MJ-
dc.date.accessioned2015-07-21T02:07:17Z-
dc.date.available2015-07-21T02:07:17Z-
dc.date.issued2015-
dc.identifier.citationPLOS ONE, 2015, v. 10 n. 6, p. e0130670-
dc.identifier.issn1932-6203-
dc.identifier.urihttp://hdl.handle.net/10722/211677-
dc.description.abstractWhen located in the endoplasmic reticulum (ER) membrane, B-cell receptor associated protein 31 (BAP31) is involved in the export of secreted proteins from the ER to the plasma membrane. In a previous study, we generated two monoclonal antibodies (mAbs), 297-D4 and 144-A8, that bound to surface molecules on human embryonic stem cells (hESCs), but not to surface molecules on mouse embryonic stem cells (mESCs). Subsequent studies revealed that the mAbs recognized BAP31 on the surface of hESCs. To investigate the membrane topology of BAP31 on the cell surface, we first examined the epitope specificity of 297-D4 and 144-A8, as well as a polyclonal anti-BAP31 antibody (alpha-BAP31). We generated a series of GST-fused BAP31 mutant proteins in which BAP31 was serially deleted at the C-terminus. GST-fused BAP31 mutant proteins were then screened to identify the epitopes targeted by the antibodies. Both 297-D4 and 144-A8 recognized C-terminal residues 208-217, while alpha-BAP31 recognized C-terminal residues 165-246, of BAP31 on hESCs, suggesting that the C-terminal domain of BAP31 is exposed on the cell surface. The polyclonal antibody alpha-BAP31 bound to mESCs, which confirmed that the C-terminal domain of BAP31 is also exposed on the surface of these cells. Our results show for the first time the novel membrane topology of cell surface-expressed BAP31 as the extracellular exposure of the BAP31 C-terminal domain was not predicted from previous studies.-
dc.languageeng-
dc.relation.ispartofPLOS ONE-
dc.rightsCreative Commons: Attribution 3.0 Hong Kong License-
dc.titleEpitope Mapping of Antibodies Suggests the Novel Membrane Topology of B-Cell Receptor Associated Protein 31 on the Cell Surface of Embryonic Stem Cells: The Novel Membrane Topology of BAP31-
dc.typeArticle-
dc.identifier.emailJung, HS: hsjung@hku.hk-
dc.identifier.authorityJung, HS=rp01683-
dc.description.naturepublished_or_final_version-
dc.identifier.doi10.1371/journal.pone.0130670-
dc.identifier.pmid26102500-
dc.identifier.hkuros244742-
dc.identifier.volume10-
dc.identifier.spagee0130670-
dc.identifier.epagee0130670-
dc.identifier.isiWOS:000356901900055-

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