title: Glycosylation is crucial for a proper catalytic site organization in human glucocerebrosidase.
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.
authors.affiliation: Centro de Biotecnologia, Universidade Federal do Rio Grande do Sul, Avenida Bento Gonçalves 9500, P.O. BOX 15005, Porto Alegre, RS, 91500-970, Brazil. laerciofachin@cbiot.ufrgs.br.|Departamento de Química Fundamental, Universidade Federal de Pernambuco, Avenida Prof. Luiz Freire, s/n, Cidade Universitária, Recife, PE, 50740-540, Brazil. laerciofachin@cbiot.ufrgs.br.
authors.forename: Laercio
authors.identifier: http://orcid.org/0000-0002-4621-3031
authors.initials: L
authors.lastname: Pol-Fachin
authors.affiliation: Centro de Biotecnologia, Universidade Federal do Rio Grande do Sul, Avenida Bento Gonçalves 9500, P.O. BOX 15005, Porto Alegre, RS, 91500-970, Brazil.|Serviço de Genética Médica, Hospital de Clínicas de Porto Alegre, Rua Ramiro Barcelos 2350, Porto Alegre, RS, 90035-903, Brazil.
authors.forename: Marina
authors.initials: M
authors.lastname: Siebert
authors.affiliation: Centro de Biotecnologia, Universidade Federal do Rio Grande do Sul, Avenida Bento Gonçalves 9500, P.O. BOX 15005, Porto Alegre, RS, 91500-970, Brazil.
authors.forename: Hugo
authors.initials: H
authors.lastname: Verli
authors.affiliation: Centro de Biotecnologia, Universidade Federal do Rio Grande do Sul, Avenida Bento Gonçalves 9500, P.O. BOX 15005, Porto Alegre, RS, 91500-970, Brazil.|Serviço de Genética Médica, Hospital de Clínicas de Porto Alegre, Rua Ramiro Barcelos 2350, Porto Alegre, RS, 90035-903, Brazil.|Departamento de Bioquímica, Universidade Federal do Rio Grande do Sul, Rua Ramiro Barcelos 2600, Porto Alegre, RS, 90035-903, Brazil.
authors.forename: Maria Luiza
authors.initials: ML
authors.lastname: Saraiva-Pereira
mesh_terms: D020134:Catalytic Domain; D005776:Gaucher Disease / Q000201:enzymology; D005962:Glucosylceramidase / Q000737:chemistry*; D006031:Glycosylation; D006801:Humans; D056004:Molecular Dynamics Simulation
chemical_list: D005962:Glucosylceramidase
keywords: Enzyme; Gaucher disease; Glycan; Lysosome; Molecular dynamics