Dimerization of the glucan phosphatase laforin requires the participation of cysteine 329.

Laforin, encoded by a gene that is mutated in Lafora Disease (LD, OMIM 254780), is a modular protein composed of a carbohydrate-binding module and a dual-specificity phosphatase domain. Laforin is the founding member of the glucan-phosphatase family and regulates the levels of phosphate present in g...

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Autores principales:	Pablo Sánchez-Martín, Madushi Raththagala, Travis M Bridges, Satrio Husodo, Matthew S Gentry, Pascual Sanz, Carlos Romá-Mateo
Formato:	article
Lenguaje:	EN
Publicado:	Public Library of Science (PLoS) 2013
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/7122f49752f34a8eb9688f8e4986fdbc
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spelling	oai:doaj.org-article:7122f49752f34a8eb9688f8e4986fdbc2021-11-18T09:02:45ZDimerization of the glucan phosphatase laforin requires the participation of cysteine 329.1932-620310.1371/journal.pone.0069523https://doaj.org/article/7122f49752f34a8eb9688f8e4986fdbc2013-01-01T00:00:00Zhttps://www.ncbi.nlm.nih.gov/pmc/articles/pmid/23922729/?tool=EBIhttps://doaj.org/toc/1932-6203Laforin, encoded by a gene that is mutated in Lafora Disease (LD, OMIM 254780), is a modular protein composed of a carbohydrate-binding module and a dual-specificity phosphatase domain. Laforin is the founding member of the glucan-phosphatase family and regulates the levels of phosphate present in glycogen. Multiple reports have described the capability of laforin to form dimers, although the function of these dimers and their relationship with LD remains unclear. Recent evidence suggests that laforin dimerization depends on redox conditions, suggesting that disulfide bonds are involved in laforin dimerization. Using site-directed mutagenesis we constructed laforin mutants in which individual cysteine residues were replaced by serine and then tested the ability of each protein to dimerize using recombinant protein as well as a mammalian cell culture assay. Laforin-Cys329Ser was the only Cys/Ser mutant unable to form dimers in both assays. We also generated a laforin truncation lacking the last three amino acids, laforin-Cys329X, and this truncation also failed to dimerize. Interestingly, laforin-Cys329Ser and laforin-Cys329X were able to bind glucans, and maintained wild type phosphatase activity against both exogenous and biologically relevant substrates. Furthermore, laforin-Cys329Ser was fully capable of participating in the ubiquitination process driven by a laforin-malin complex. These results suggest that dimerization is not required for laforin phosphatase activity, glucan binding, or for the formation of a functional laforin-malin complex. Cumulatively, these results suggest that cysteine 329 is specifically involved in the dimerization process of laforin. Therefore, the C329S mutant constitutes a valuable tool to analyze the physiological implications of laforin's oligomerization.Pablo Sánchez-MartínMadushi RaththagalaTravis M BridgesSatrio HusodoMatthew S GentryPascual SanzCarlos Romá-MateoPublic Library of Science (PLoS)articleMedicineRScienceQENPLoS ONE, Vol 8, Iss 7, p e69523 (2013)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Pablo Sánchez-Martín Madushi Raththagala Travis M Bridges Satrio Husodo Matthew S Gentry Pascual Sanz Carlos Romá-Mateo Dimerization of the glucan phosphatase laforin requires the participation of cysteine 329.
description	Laforin, encoded by a gene that is mutated in Lafora Disease (LD, OMIM 254780), is a modular protein composed of a carbohydrate-binding module and a dual-specificity phosphatase domain. Laforin is the founding member of the glucan-phosphatase family and regulates the levels of phosphate present in glycogen. Multiple reports have described the capability of laforin to form dimers, although the function of these dimers and their relationship with LD remains unclear. Recent evidence suggests that laforin dimerization depends on redox conditions, suggesting that disulfide bonds are involved in laforin dimerization. Using site-directed mutagenesis we constructed laforin mutants in which individual cysteine residues were replaced by serine and then tested the ability of each protein to dimerize using recombinant protein as well as a mammalian cell culture assay. Laforin-Cys329Ser was the only Cys/Ser mutant unable to form dimers in both assays. We also generated a laforin truncation lacking the last three amino acids, laforin-Cys329X, and this truncation also failed to dimerize. Interestingly, laforin-Cys329Ser and laforin-Cys329X were able to bind glucans, and maintained wild type phosphatase activity against both exogenous and biologically relevant substrates. Furthermore, laforin-Cys329Ser was fully capable of participating in the ubiquitination process driven by a laforin-malin complex. These results suggest that dimerization is not required for laforin phosphatase activity, glucan binding, or for the formation of a functional laforin-malin complex. Cumulatively, these results suggest that cysteine 329 is specifically involved in the dimerization process of laforin. Therefore, the C329S mutant constitutes a valuable tool to analyze the physiological implications of laforin's oligomerization.
format	article
author	Pablo Sánchez-Martín Madushi Raththagala Travis M Bridges Satrio Husodo Matthew S Gentry Pascual Sanz Carlos Romá-Mateo
author_facet	Pablo Sánchez-Martín Madushi Raththagala Travis M Bridges Satrio Husodo Matthew S Gentry Pascual Sanz Carlos Romá-Mateo
author_sort	Pablo Sánchez-Martín
title	Dimerization of the glucan phosphatase laforin requires the participation of cysteine 329.
title_short	Dimerization of the glucan phosphatase laforin requires the participation of cysteine 329.
title_full	Dimerization of the glucan phosphatase laforin requires the participation of cysteine 329.
title_fullStr	Dimerization of the glucan phosphatase laforin requires the participation of cysteine 329.
title_full_unstemmed	Dimerization of the glucan phosphatase laforin requires the participation of cysteine 329.
title_sort	dimerization of the glucan phosphatase laforin requires the participation of cysteine 329.
publisher	Public Library of Science (PLoS)
publishDate	2013
url	https://doaj.org/article/7122f49752f34a8eb9688f8e4986fdbc
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Dimerization of the glucan phosphatase laforin requires the participation of cysteine 329.

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