Cargando…

Glycosylation of Erythrocyte Spectrin and Its Modification in Visceral Leishmaniasis

Using a lectin, Achatinin-H, having preferential specificity for glycoproteins with terminal 9-O-acetyl sialic acid derivatives linked in α2-6 linkages to subterminal N-acetylgalactosamine, eight distinct disease-associated 9-O-acetylated sialoglycoproteins was purified from erythrocytes of visceral...

Descripción completa

Detalles Bibliográficos
Autores principales: Samanta, Sajal, Dutta, Devawati, Ghoshal, Angana, Mukhopadhyay, Sumi, Saha, Bibhuti, Sundar, Shyam, Jarmalavicius, Saulius, Forgber, Michael, Mandal, Chhabinath, Walden, Peter, Mandal, Chitra
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Public Library of Science 2011
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3229537/
https://www.ncbi.nlm.nih.gov/pubmed/22164239
http://dx.doi.org/10.1371/journal.pone.0028169
_version_ 1782217959045857280
author Samanta, Sajal
Dutta, Devawati
Ghoshal, Angana
Mukhopadhyay, Sumi
Saha, Bibhuti
Sundar, Shyam
Jarmalavicius, Saulius
Forgber, Michael
Mandal, Chhabinath
Walden, Peter
Mandal, Chitra
author_facet Samanta, Sajal
Dutta, Devawati
Ghoshal, Angana
Mukhopadhyay, Sumi
Saha, Bibhuti
Sundar, Shyam
Jarmalavicius, Saulius
Forgber, Michael
Mandal, Chhabinath
Walden, Peter
Mandal, Chitra
author_sort Samanta, Sajal
collection PubMed
description Using a lectin, Achatinin-H, having preferential specificity for glycoproteins with terminal 9-O-acetyl sialic acid derivatives linked in α2-6 linkages to subterminal N-acetylgalactosamine, eight distinct disease-associated 9-O-acetylated sialoglycoproteins was purified from erythrocytes of visceral leishmaniaisis (VL) patients (RBC(VL)). Analyses of tryptic fragments by mass spectrometry led to the identification of two high-molecular weight 9-O-acetylated sialoglycoproteins as human erythrocytic α- and β-spectrin. Total spectrin purified from erythrocytes of VL patients (spectrin(VL)) was reactive with Achatinin-H. Interestingly, along with two high molecular weight bands corresponding to α- and β-spectrin another low molecular weight 60 kDa band was observed. Total spectrin was also purified from normal human erythrocytes (spectrin(N)) and insignificant binding with Achatinin-H was demonstrated. Additionally, this 60 kDa fragment was totally absent in spectrin(N). Although the presence of both N- and O-glycosylations was found both in spectrin(N) and spectrin(VL), enhanced sialylation was predominantly induced in spectrin(VL). Sialic acids accounted for approximately 1.25 kDa mass of the 60 kDa polypeptide. The demonstration of a few identified sialylated tryptic fragments of α- and β-spectrin(VL) confirmed the presence of terminal sialic acids. Molecular modelling studies of spectrin suggest that a sugar moiety can fit into the potential glycosylation sites. Interestingly, highly sialylated spectrin(VL) showed decreased binding with spectrin-depleted inside-out membrane vesicles of normal erythrocytes compared to spectrin(N) suggesting functional abnormality. Taken together this is the first report of glycosylated eythrocytic spectrin in normal erythrocytes and its enhanced sialylation in RBC(VL). The enhanced sialylation of this cytoskeleton protein is possibly related to the fragmentation of spectrin(VL) as evidenced by the presence of an additional 60 kDa fragment, absent in spectrin(N) which possibly affects the biology of RBC(VL) linked to both severe distortion of erythrocyte development and impairment of erythrocyte membrane integrity and may provide an explanation for their sensitivity to hemolysis and anemia in VL patients.
format Online
Article
Text
id pubmed-3229537
institution National Center for Biotechnology Information
language English
publishDate 2011
publisher Public Library of Science
record_format MEDLINE/PubMed
spelling pubmed-32295372011-12-07 Glycosylation of Erythrocyte Spectrin and Its Modification in Visceral Leishmaniasis Samanta, Sajal Dutta, Devawati Ghoshal, Angana Mukhopadhyay, Sumi Saha, Bibhuti Sundar, Shyam Jarmalavicius, Saulius Forgber, Michael Mandal, Chhabinath Walden, Peter Mandal, Chitra PLoS One Research Article Using a lectin, Achatinin-H, having preferential specificity for glycoproteins with terminal 9-O-acetyl sialic acid derivatives linked in α2-6 linkages to subterminal N-acetylgalactosamine, eight distinct disease-associated 9-O-acetylated sialoglycoproteins was purified from erythrocytes of visceral leishmaniaisis (VL) patients (RBC(VL)). Analyses of tryptic fragments by mass spectrometry led to the identification of two high-molecular weight 9-O-acetylated sialoglycoproteins as human erythrocytic α- and β-spectrin. Total spectrin purified from erythrocytes of VL patients (spectrin(VL)) was reactive with Achatinin-H. Interestingly, along with two high molecular weight bands corresponding to α- and β-spectrin another low molecular weight 60 kDa band was observed. Total spectrin was also purified from normal human erythrocytes (spectrin(N)) and insignificant binding with Achatinin-H was demonstrated. Additionally, this 60 kDa fragment was totally absent in spectrin(N). Although the presence of both N- and O-glycosylations was found both in spectrin(N) and spectrin(VL), enhanced sialylation was predominantly induced in spectrin(VL). Sialic acids accounted for approximately 1.25 kDa mass of the 60 kDa polypeptide. The demonstration of a few identified sialylated tryptic fragments of α- and β-spectrin(VL) confirmed the presence of terminal sialic acids. Molecular modelling studies of spectrin suggest that a sugar moiety can fit into the potential glycosylation sites. Interestingly, highly sialylated spectrin(VL) showed decreased binding with spectrin-depleted inside-out membrane vesicles of normal erythrocytes compared to spectrin(N) suggesting functional abnormality. Taken together this is the first report of glycosylated eythrocytic spectrin in normal erythrocytes and its enhanced sialylation in RBC(VL). The enhanced sialylation of this cytoskeleton protein is possibly related to the fragmentation of spectrin(VL) as evidenced by the presence of an additional 60 kDa fragment, absent in spectrin(N) which possibly affects the biology of RBC(VL) linked to both severe distortion of erythrocyte development and impairment of erythrocyte membrane integrity and may provide an explanation for their sensitivity to hemolysis and anemia in VL patients. Public Library of Science 2011-12-02 /pmc/articles/PMC3229537/ /pubmed/22164239 http://dx.doi.org/10.1371/journal.pone.0028169 Text en Samanta et al. http://creativecommons.org/licenses/by/4.0/ 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 properly credited.
spellingShingle Research Article
Samanta, Sajal
Dutta, Devawati
Ghoshal, Angana
Mukhopadhyay, Sumi
Saha, Bibhuti
Sundar, Shyam
Jarmalavicius, Saulius
Forgber, Michael
Mandal, Chhabinath
Walden, Peter
Mandal, Chitra
Glycosylation of Erythrocyte Spectrin and Its Modification in Visceral Leishmaniasis
title Glycosylation of Erythrocyte Spectrin and Its Modification in Visceral Leishmaniasis
title_full Glycosylation of Erythrocyte Spectrin and Its Modification in Visceral Leishmaniasis
title_fullStr Glycosylation of Erythrocyte Spectrin and Its Modification in Visceral Leishmaniasis
title_full_unstemmed Glycosylation of Erythrocyte Spectrin and Its Modification in Visceral Leishmaniasis
title_short Glycosylation of Erythrocyte Spectrin and Its Modification in Visceral Leishmaniasis
title_sort glycosylation of erythrocyte spectrin and its modification in visceral leishmaniasis
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3229537/
https://www.ncbi.nlm.nih.gov/pubmed/22164239
http://dx.doi.org/10.1371/journal.pone.0028169
work_keys_str_mv AT samantasajal glycosylationoferythrocytespectrinanditsmodificationinvisceralleishmaniasis
AT duttadevawati glycosylationoferythrocytespectrinanditsmodificationinvisceralleishmaniasis
AT ghoshalangana glycosylationoferythrocytespectrinanditsmodificationinvisceralleishmaniasis
AT mukhopadhyaysumi glycosylationoferythrocytespectrinanditsmodificationinvisceralleishmaniasis
AT sahabibhuti glycosylationoferythrocytespectrinanditsmodificationinvisceralleishmaniasis
AT sundarshyam glycosylationoferythrocytespectrinanditsmodificationinvisceralleishmaniasis
AT jarmalaviciussaulius glycosylationoferythrocytespectrinanditsmodificationinvisceralleishmaniasis
AT forgbermichael glycosylationoferythrocytespectrinanditsmodificationinvisceralleishmaniasis
AT mandalchhabinath glycosylationoferythrocytespectrinanditsmodificationinvisceralleishmaniasis
AT waldenpeter glycosylationoferythrocytespectrinanditsmodificationinvisceralleishmaniasis
AT mandalchitra glycosylationoferythrocytespectrinanditsmodificationinvisceralleishmaniasis