Anaerobic sulfatase maturase AslB from Escherichia coli activates human recombinant iduronate-2-sulfate sulfatase (IDS) and N-acetylgalactosamine-6-sulfate sulfatase (GALNS)

Maturation of type I sulfatases requires the conversion of the cysteine (Cys) or serine (Ser) present in the active site to formylglycine (FGly). This activation represents a limiting step during the production of recombinant sulfatases in bacteria and eukaryotic hosts. AslB, YdeM and YidF have been proposed to participate in the activation of sulfatases in Escherichia coli. In this study, we combined in-silico and experimental approaches to study the interaction between Escherichia coli BL21(DE3) AslB and human sulfatases, more specifically iduronate-2-sulfate sulfatase (IDS) and N-acetylgalactosamine-6-sulfate sulfatase (GALNS). In-silico results show that AslB has a higher affinity for the residual motif of GALNS (? 9.4 kcal mol? 1), Cys- and Ser-type, than for the one of IDS (? 8.0 kcal mol? 1). However, the distance between the AslB active residue and the target motif favors the interaction with IDS (4.4 Å) more than with GALNS (5.5 Å). Experimental observations supported in-silico results where the co-expression of AslB with GALNS Cys- and Ser-type presented an activity increment of 2.0- and 1.5-fold compared to the control cultures, lacking overexpressed AslB. Similarly, IDS activity was increased in 4.6-fold when co-expressed with AslB. The higher sulfatase activity of AslB-IDS suggests that the distance between the AslB active residue and the motif target is a key parameter for the in-silico search of potential sulfatase activators. In conclusion, our results suggest that AslB is involve in the maturation of heterologous human sulfatases in E. coli BL21(DE3), and that it can have important implications in the production of recombinant sulfatases for therapeutic purposes and research. © 2017 Elsevier B.V.

• Fecha de publicación: 2017
• Institución: Universidad del Rosario
• Repositorio: Repositorio EdocUR - U. Rosario
• Idioma: eng
• OAI Identifier: oai:repository.urosario.edu.co:10336/24133
• Acceso en línea: https://doi.org/10.1016/j.gene.2017.08.043; https://repository.urosario.edu.co/handle/10336/24133
• Palabra clave: Anaerobic sulfatase maturase aslb; Cysteine; Iduronate 2 sulfatase; N acetylgalactosamine 6 sulfatase; Recombinant enzyme; Serine; Sulfatase; Unclassified drug; Escherichia coli protein; Glycoprotein; N acetylgalactosamine 4 sulfatase; Protein binding; Recombinant protein; Sulfatase; Amino acid sequence; Article; Binding affinity; Controlled study; Enzyme activation; Enzyme active site; Enzyme activity; Escherichia coli; Molecular docking; Molecular dynamics; Molecular model; Nonhuman; Priority journal; Protein expression; Protein motif; Protein protein interaction; Chemistry; Enzyme activation; Enzymology; Escherichia coli; Human; Metabolism; Catalytic Domain; Chondroitinsulfatases; Cysteine; Enzyme Activation; Escherichia coli; Escherichia coli Proteins; Glycoproteins; Humans; Molecular Docking Simulation; Molecular Dynamics Simulation; Protein Binding; Recombinant Proteins; Serine; Sulfatases; Aslb; Formylglycine-generating enzymes; Iduronate-2-sulfate sulfatase; N-acetylgalactosamine-6-sulfate sulfatase; Sulfatases; human; human; Molecular; GALNS protein; IDS protein; Models
• License: Abierto (Texto Completo)