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Glycosylation is crucial for a proper catalytic site organization in human glucocerebrosidase

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Abstract

Gaucher disease, an autosomal recessive disorder, is caused by a deficiency of glucocerebrosidase (GCase) enzyme, a peripheral membrane-associated glycoprotein that hydrolyses glucosylceramide in lysosomes. Glycosylation is essential for the development of a catalytically active enzyme, specifically in the first site, located at Asn19. However, both the molecular basis of the relevance of N-glycosylation over GCase activity and the effects of glycosylation over its structure and dynamics are still not fully understood. Thus, the present work evaluated GCase enzyme in increasing glycosylation content using triplicate unbiased molecular dynamics simulations. Accordingly, the N-linked glycan chains caused local conformational stabilization effects over the protein, as well as in regions flanking the enzyme catalytic dyad. In the case of the Asn19-linked glycan, it also occurred around region 438–444, where one of the most prevalent GCase mutations is found. Markedly, an increasing catalytic dyad organization was related to increasing glycosylation contents, offering the first atomic-level explanation for the experimental observation that GCase activity is controlled by glycosylation, especially at Asn19.

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Authors and Affiliations

  1. Centro de Biotecnologia, Universidade Federal do Rio Grande do Sul, Avenida Bento Gonçalves 9500, P.O. BOX 15005, Porto Alegre, RS, 91500-970, Brazil

    Laercio Pol-Fachin, Marina Siebert, Hugo Verli & Maria Luiza Saraiva-Pereira

  2. Departamento de Química Fundamental, Universidade Federal de Pernambuco, Avenida Prof. Luiz Freire, s/n, Cidade Universitária, Recife, PE, 50740-540, Brazil

    Laercio Pol-Fachin

  3. Serviço de Genética Médica, Hospital de Clínicas de Porto Alegre, Rua Ramiro Barcelos 2350, Porto Alegre, RS, 90035-903, Brazil

    Marina Siebert & Maria Luiza Saraiva-Pereira

  4. Departamento de Bioquímica, Universidade Federal do Rio Grande do Sul, Rua Ramiro Barcelos 2600, Porto Alegre, RS, 90035-903, Brazil

    Maria Luiza Saraiva-Pereira

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  1. Laercio Pol-Fachin
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Correspondence to Laercio Pol-Fachin.

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This work was supported by Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq – MCT, DCR-0037-1.06/13); Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES – MEC); Fundação de Amparo à Pesquisa do Rio Grande do Sul (FAPERGS); Fundação de Amparo a Ciência e Tecnologia do Estado de Pernambuco (FACEPE, grant number APQ-0398-1.06/13), and Fundo de Incentivo à Pesquisa e Eventos do Hospital de Clínicas de Porto Alegre (FIPE-HCPA).

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Pol-Fachin, L., Siebert, M., Verli, H. et al. Glycosylation is crucial for a proper catalytic site organization in human glucocerebrosidase. Glycoconj J 33, 237–244 (2016). https://doi.org/10.1007/s10719-016-9661-7

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  • DOI: https://doi.org/10.1007/s10719-016-9661-7

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