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(Referência obtida automaticamente do Web of Science, por meio da informação sobre o financiamento pela FAPESP e o número do processo correspondente, incluída na publicação pelos autores.)

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

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Autor(es):
Gesteira, Tarsis F. [1] ; Pol-Fachin, Laercio [2] ; Coulson-Thomas, Vivien Jane [1] ; Lima, Marcelo A. [1] ; Verli, Hugo [2] ; Nader, Helena B. [1]
Número total de Autores: 6
Afiliação do(s) autor(es):
[1] Univ Fed Sao Paulo, Dept Bioquim, Sao Paulo - Brazil
[2] Univ Fed Rio Grande do Sul, Ctr Biotecnol, Porto Alegre, RS - Brazil
Número total de Afiliações: 2
Tipo de documento: Artigo Científico
Fonte: PLoS One; v. 8, n. 8 AUG 5 2013.
Citações Web of Science: 10
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. (AU)

Processo FAPESP: 10/52426-3 - Espectrometria de massas e de ressonância magnética nuclear na caracterização estrutural de glicosaminoglicanos e polissacarídeos complexos de invertebrados e algas
Beneficiário:Helena Bonciani Nader
Linha de fomento: Auxílio à Pesquisa - Regular