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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...

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Autores principales: Karbian, Netanel, Eshed-Eisenbach, Yael, Tabib, Adi, Hoizman, Hila, Morgan, B. Paul, Schueler-Furman, Ora, Peles, Elior, Mevorach, Dror
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Wolters Kluwer 2018
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.
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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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