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      Correction: 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

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          Abstract

          In Fig 5A, the N-terminal residue of the underlined epitope is incorrectly listed as Glutamate (E) instead of Glutamine (Q). Please see the corrected Fig 5 here. 10.1371/journal.pone.0170145.g001 Fig 5 BAP31 is also expressed on the surface of mESCs. (A) The epitope sequences of 297-D4 and 144-A8 are different between hESCs and mESCs. The partial C-terminal amino acid sequences were aligned between human and mouse BAP31. The epitope sequences are underlined, and the different amino acids are indicated by arrows. (B) Flow cytometry analysis of mESCs with anti-SSEA-1, 297-D4, 144-A8, and α-BAP31 antibodies. (C) Immunoprecipitation of mESCs with 297-D4, 144-A8, and α-BAP31 antibodies. Cell surface proteins of mESCs were biotinylated, immuoprecipitated, and analyzed by SA-HRP or western blot with α-BAP31 (left panels). The input samples were also analyzed by western blots (right panels). β-actin was used as a loading control.

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          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

          When 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 (α-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 α-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 α-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.
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            Author and article information

            Journal
            PLoS One
            PLoS ONE
            plos
            plosone
            PLoS ONE
            Public Library of Science (San Francisco, CA USA )
            1932-6203
            6 January 2017
            2017
            : 12
            : 1
            : e0170145
            Article
            PONE-D-16-51568
            10.1371/journal.pone.0170145
            5218809
            28061503
            16e0dea0-6698-4817-b0e1-fd7d9a455170
            © 2017 Kim et al

            This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.

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