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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...

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Autores principales: Matalkah, Fatimah, Rhodes, Scott, Ramamurthy, Visvanathan, Stoilov, Peter
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Public Library of Science 2022
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.
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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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