Insights into the N-Sulfation Mechanism: Molecular Dynamics Simulations of the N-Sulfotransferase Domain of Ndst1 and Mutants

Insights into the N-Sulfation Mechanism: Molecular Dynamics Simulations of the N-Sulfotransferase Domain of Ndst1 and Mutants

Autor Gesteira, Tarsis F. Autor UNIFESP Google Scholar
Pol-Fachin, Laercio Google Scholar
Coulson-Thomas, Vivien Jane Autor UNIFESP Google Scholar
Lima, Marcelo A. Autor UNIFESP Google Scholar
Verli, Hugo Google Scholar
Nader, Helena Bonciani Autor UNIFESP Google Scholar
Instituição Universidade Federal de São Paulo (UNIFESP)
Univ Fed Rio Grande do Sul
Resumo Sulfation patterns along glycosaminoglycan (GAG) chains dictate their functional role. the N-deacetylase N-sulfotransferase family (NDST) catalyzes the initial downstream modification of heparan sulfate and heparin chains by removing acetyl groups from subsets of N-acetylglucosamine units and, subsequently, sulfating the residual free amino groups. These enzymes transfer the sulfuryl group from 3'-phosphoadenosine-5'-phosphosulfate (PAPS), yielding sulfated sugar chains and 3'-phosphoadenosine-5'-phosphate (PAP). for the N-sulfotransferase domain of NDST1, Lys833 has been implicated to play a role in holding the substrate glycan moiety close to the PAPS cofactor. Additionally, Lys833 together with His716 interact with the sulfonate group, stabilizing the transition state. Such a role seems to be shared by Lys614 through donation of a proton to the bridging oxygen of the cofactor, thereby acting as a catalytic acid. However, the relevance of these boundary residues at the hydrophobic cleft is still unclear. Moreover, whether Lys833, His716 and Lys614 play a role in both glycan recognition and glycan sulfation remains elusive. in this study we evaluate the contribution of NDST mutants (Lys833, His716 and Lys614) to dynamical effects during sulfate transfer using comprehensive combined docking and essential dynamics. in addition, the binding location of the glycan moiety, PAPS and PAP within the active site of NDST1 throughout the sulfate transfer were determined by intermediate state analysis. Furthermore, NDST1 mutants unveiled Lys833 as vital for both the glycan binding and subsequent N-sulfotransferase activity of NDST1.
Idioma Inglês
Financiador Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)
Conselho Nacional do Desenvolvimento Cientifico e Tecnologico
Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)
Número do financiamento FAPESP: 2010/52426-3
Data 2013-08-05
Publicado em Plos One. San Francisco: Public Library Science, v. 8, n. 8, 12 p., 2013.
ISSN 1932-6203 (Sherpa/Romeo, fator de impacto)
Editor Public Library Science
Extensão 12
Fonte http://dx.doi.org/10.1371/journal.pone.0070880
Direito de acesso Acesso aberto Open Access
Tipo Artigo
Web of Science WOS:000324465000147
URI http://repositorio.unifesp.br/handle/11600/36643

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