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Molecular pathogenesis of human CD59 deficiency
OBJECTIVE: To characterize all 4 mutations described for CD59 congenital deficiency. METHODS: The 4 mutations, p.Cys64Tyr, p.Asp24Val, p.Asp24Valfs*, and p.Ala16Alafs*, were described in 13 individuals with CD59 malfunction. All 13 presented with recurrent Guillain-Barré syndrome or chronic inflamma...
Autores principales: | , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Wolters Kluwer
2018
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6244018/ https://www.ncbi.nlm.nih.gov/pubmed/30533526 http://dx.doi.org/10.1212/NXG.0000000000000280 |
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author | Karbian, Netanel Eshed-Eisenbach, Yael Tabib, Adi Hoizman, Hila Morgan, B. Paul Schueler-Furman, Ora Peles, Elior Mevorach, Dror |
author_facet | Karbian, Netanel Eshed-Eisenbach, Yael Tabib, Adi Hoizman, Hila Morgan, B. Paul Schueler-Furman, Ora Peles, Elior Mevorach, Dror |
author_sort | Karbian, Netanel |
collection | PubMed |
description | OBJECTIVE: To characterize all 4 mutations described for CD59 congenital deficiency. METHODS: The 4 mutations, p.Cys64Tyr, p.Asp24Val, p.Asp24Valfs*, and p.Ala16Alafs*, were described in 13 individuals with CD59 malfunction. All 13 presented with recurrent Guillain-Barré syndrome or chronic inflammatory demyelinating polyneuropathy, recurrent strokes, and chronic hemolysis. Here, we track the molecular consequences of the 4 mutations and their effects on CD59 expression, localization, glycosylation, degradation, secretion, and function. Mutants were cloned and inserted into plasmids to analyze their expression, localization, and functionality. RESULTS: Immunolabeling of myc-tagged wild-type (WT) and mutant CD59 proteins revealed cell surface expression of p.Cys64Tyr and p.Asp24Val detected with the myc antibody, but no labeling by anti-CD59 antibodies. In contrast, frameshift mutants p.Asp24Valfs* and p.Ala16Alafs* were detected only intracellularly and did not reach the cell surface. Western blot analysis showed normal glycosylation but mutant-specific secretion patterns. All mutants significantly increased MAC-dependent cell lysis compared with WT. In contrast to CD59 knockout mice previously used to characterize phenotypic effects of CD59 perturbation, all 4 hCD59 mutations generate CD59 proteins that are expressed and may function intracellularly (4) or on the cell membrane (2). None of the 4 CD59 mutants are detected by known anti-CD59 antibodies, including the 2 variants present on the cell membrane. None of the 4 inhibits membrane attack complex (MAC) formation. CONCLUSIONS: All 4 mutants generate nonfunctional CD59, 2 are expressed as cell surface proteins that may function in non–MAC-related interactions and 2 are expressed only intracellularly. Distinct secretion of soluble CD59 may have also a role in disease pathogenesis. |
format | Online Article Text |
id | pubmed-6244018 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2018 |
publisher | Wolters Kluwer |
record_format | MEDLINE/PubMed |
spelling | pubmed-62440182018-12-07 Molecular pathogenesis of human CD59 deficiency Karbian, Netanel Eshed-Eisenbach, Yael Tabib, Adi Hoizman, Hila Morgan, B. Paul Schueler-Furman, Ora Peles, Elior Mevorach, Dror Neurol Genet Article OBJECTIVE: To characterize all 4 mutations described for CD59 congenital deficiency. METHODS: The 4 mutations, p.Cys64Tyr, p.Asp24Val, p.Asp24Valfs*, and p.Ala16Alafs*, were described in 13 individuals with CD59 malfunction. All 13 presented with recurrent Guillain-Barré syndrome or chronic inflammatory demyelinating polyneuropathy, recurrent strokes, and chronic hemolysis. Here, we track the molecular consequences of the 4 mutations and their effects on CD59 expression, localization, glycosylation, degradation, secretion, and function. Mutants were cloned and inserted into plasmids to analyze their expression, localization, and functionality. RESULTS: Immunolabeling of myc-tagged wild-type (WT) and mutant CD59 proteins revealed cell surface expression of p.Cys64Tyr and p.Asp24Val detected with the myc antibody, but no labeling by anti-CD59 antibodies. In contrast, frameshift mutants p.Asp24Valfs* and p.Ala16Alafs* were detected only intracellularly and did not reach the cell surface. Western blot analysis showed normal glycosylation but mutant-specific secretion patterns. All mutants significantly increased MAC-dependent cell lysis compared with WT. In contrast to CD59 knockout mice previously used to characterize phenotypic effects of CD59 perturbation, all 4 hCD59 mutations generate CD59 proteins that are expressed and may function intracellularly (4) or on the cell membrane (2). None of the 4 CD59 mutants are detected by known anti-CD59 antibodies, including the 2 variants present on the cell membrane. None of the 4 inhibits membrane attack complex (MAC) formation. CONCLUSIONS: All 4 mutants generate nonfunctional CD59, 2 are expressed as cell surface proteins that may function in non–MAC-related interactions and 2 are expressed only intracellularly. Distinct secretion of soluble CD59 may have also a role in disease pathogenesis. Wolters Kluwer 2018-10-26 /pmc/articles/PMC6244018/ /pubmed/30533526 http://dx.doi.org/10.1212/NXG.0000000000000280 Text en Copyright © 2018 The Author(s). Published by Wolters Kluwer Health, Inc. on behalf of the American Academy of Neurology. This is an open access article distributed under the terms of the Creative Commons Attribution-NonCommercial-NoDerivatives License 4.0 (CC BY-NC-ND) (http://creativecommons.org/licenses/by-nc-nd/4.0/) , which permits downloading and sharing the work provided it is properly cited. The work cannot be changed in any way or used commercially without permission from the journal. |
spellingShingle | Article Karbian, Netanel Eshed-Eisenbach, Yael Tabib, Adi Hoizman, Hila Morgan, B. Paul Schueler-Furman, Ora Peles, Elior Mevorach, Dror Molecular pathogenesis of human CD59 deficiency |
title | Molecular pathogenesis of human CD59 deficiency |
title_full | Molecular pathogenesis of human CD59 deficiency |
title_fullStr | Molecular pathogenesis of human CD59 deficiency |
title_full_unstemmed | Molecular pathogenesis of human CD59 deficiency |
title_short | Molecular pathogenesis of human CD59 deficiency |
title_sort | molecular pathogenesis of human cd59 deficiency |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6244018/ https://www.ncbi.nlm.nih.gov/pubmed/30533526 http://dx.doi.org/10.1212/NXG.0000000000000280 |
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