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The mAB 13A4 monoclonal antibody to the mouse PROM1 protein recognizes a structural epitope
PROM1 (CD133, AC133) is a protein that is required for the maintenance of primary cilia. Mutation in the Prom1 gene in humans and animal models are associated with several forms of retinal degeneration. mAB 13A4 is the main reagent used to detect the mouse PROM1 protein. We endeavored to map the epi...
Autores principales: | , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Public Library of Science
2022
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9550058/ https://www.ncbi.nlm.nih.gov/pubmed/36215230 http://dx.doi.org/10.1371/journal.pone.0274958 |
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author | Matalkah, Fatimah Rhodes, Scott Ramamurthy, Visvanathan Stoilov, Peter |
author_facet | Matalkah, Fatimah Rhodes, Scott Ramamurthy, Visvanathan Stoilov, Peter |
author_sort | Matalkah, Fatimah |
collection | PubMed |
description | PROM1 (CD133, AC133) is a protein that is required for the maintenance of primary cilia. Mutation in the Prom1 gene in humans and animal models are associated with several forms of retinal degeneration. mAB 13A4 is the main reagent used to detect the mouse PROM1 protein. We endeavored to map the epitope of the rat monoclonal antibody mAB 13A4 to the mouse PROM1 protein. Deletion mutagenesis demonstrated that mAB 13A4 recognizes a structural epitope that is stabilized by two of the extracellular domains of PROM1. Furthermore, the affinity of mAB 13A4 to the major PROM1 isoform in photoreceptor cells is significantly reduced due to the inclusion of a photoreceptor-specific alternative exon in the third extracellular domain. Interestingly, a deletion in the photoreceptor specific isoform of six amino acids adjacent to the alternative exon restored the affinity of mAB 13A4 to PROM1. The results of the mutagenesis are consistent with the computationally predicted helical bundle structure of PROM1 and point to the utility of mAB 13A4 for evaluating the effect of mutations on the PROM1 structure. Our results show that the PROM1 isoform composition needs to be considered when interpreting tissue and developmental expression data produced by mAB 13A4. |
format | Online Article Text |
id | pubmed-9550058 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2022 |
publisher | Public Library of Science |
record_format | MEDLINE/PubMed |
spelling | pubmed-95500582022-10-11 The mAB 13A4 monoclonal antibody to the mouse PROM1 protein recognizes a structural epitope Matalkah, Fatimah Rhodes, Scott Ramamurthy, Visvanathan Stoilov, Peter PLoS One Research Article PROM1 (CD133, AC133) is a protein that is required for the maintenance of primary cilia. Mutation in the Prom1 gene in humans and animal models are associated with several forms of retinal degeneration. mAB 13A4 is the main reagent used to detect the mouse PROM1 protein. We endeavored to map the epitope of the rat monoclonal antibody mAB 13A4 to the mouse PROM1 protein. Deletion mutagenesis demonstrated that mAB 13A4 recognizes a structural epitope that is stabilized by two of the extracellular domains of PROM1. Furthermore, the affinity of mAB 13A4 to the major PROM1 isoform in photoreceptor cells is significantly reduced due to the inclusion of a photoreceptor-specific alternative exon in the third extracellular domain. Interestingly, a deletion in the photoreceptor specific isoform of six amino acids adjacent to the alternative exon restored the affinity of mAB 13A4 to PROM1. The results of the mutagenesis are consistent with the computationally predicted helical bundle structure of PROM1 and point to the utility of mAB 13A4 for evaluating the effect of mutations on the PROM1 structure. Our results show that the PROM1 isoform composition needs to be considered when interpreting tissue and developmental expression data produced by mAB 13A4. Public Library of Science 2022-10-10 /pmc/articles/PMC9550058/ /pubmed/36215230 http://dx.doi.org/10.1371/journal.pone.0274958 Text en © 2022 Matalkah et al https://creativecommons.org/licenses/by/4.0/This is an open access article distributed under the terms of the Creative Commons Attribution License (https://creativecommons.org/licenses/by/4.0/) , which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. |
spellingShingle | Research Article Matalkah, Fatimah Rhodes, Scott Ramamurthy, Visvanathan Stoilov, Peter The mAB 13A4 monoclonal antibody to the mouse PROM1 protein recognizes a structural epitope |
title | The mAB 13A4 monoclonal antibody to the mouse PROM1 protein recognizes a structural epitope |
title_full | The mAB 13A4 monoclonal antibody to the mouse PROM1 protein recognizes a structural epitope |
title_fullStr | The mAB 13A4 monoclonal antibody to the mouse PROM1 protein recognizes a structural epitope |
title_full_unstemmed | The mAB 13A4 monoclonal antibody to the mouse PROM1 protein recognizes a structural epitope |
title_short | The mAB 13A4 monoclonal antibody to the mouse PROM1 protein recognizes a structural epitope |
title_sort | mab 13a4 monoclonal antibody to the mouse prom1 protein recognizes a structural epitope |
topic | Research Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9550058/ https://www.ncbi.nlm.nih.gov/pubmed/36215230 http://dx.doi.org/10.1371/journal.pone.0274958 |
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