Automated Solid‐Phase Synthesis of β‐Mannuronic Acid Alginates

Walvoort, Marthe T. C.; Elst, Hans van den; Plante, Obadiah J.; Kröck, Lenz; Seeberger, Peter H.; Overkleeft, Herman S.; Marel, Gijsbert A. van der; Codée, Jeroen D. C.

doi:10.1002/anie.201108744

Cited by 100 publications

(76 citation statements)

References 30 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Each protein was concentrated and dialyzed against 50 mM ammonium acetate using microconcentrators (Millipore Corp., Bedford, MA) with a MW cut-off of 10 kDa and stored at −20°C until needed. Stock solutions of each of the polymannurnic acid oligomers, tetramer through dodecamer (ManA 4 – ManA 12 ), and undeca- and pentadeca-hyaluronic acid saccharides (HA 11 and HA 15 ), synthesized as previously described [77], [78] were prepared by dissolving known amounts of solid compound in ultrafiltered water (Milli-Q, Millipore, Bedford, MA) to give a final concentration of ∼1 mM. The ligand solutions were stored at 253 K until needed.…”

Section: Methodsmentioning

confidence: 99%

“…Briefly, 3 mM 4-nitrophenyl acetate dissolved in ethanol (3% final concentration), and 1 mM of ManA 10 [78], was incubated with 60 µg of AlgX in 50 mM sodium phosphate buffer, pH 7.0 for a duration of 1 h at 298 K. Control reactions containing 1 mM of cellohexose, xylohexose and maltotriose in place of alginate were also completed. All reactions were repeated in the absence of AlgX as a negative control.…”

Section: Methodsmentioning

confidence: 99%

See 1 more Smart Citation

P. aeruginosa SGNH Hydrolase-Like Proteins AlgJ and AlgX Have Similar Topology but Separate and Distinct Roles in Alginate Acetylation

et al. 2014

Self Cite

View full text Add to dashboard Cite

The O-acetylation of polysaccharides is a common modification used by pathogenic organisms to protect against external forces. Pseudomonas aeruginosa secretes the anionic, O-acetylated exopolysaccharide alginate during chronic infection in the lungs of cystic fibrosis patients to form the major constituent of a protective biofilm matrix. Four proteins have been implicated in the O-acetylation of alginate, AlgIJF and AlgX. To probe the biological function of AlgJ, we determined its structure to 1.83 Å resolution. AlgJ is a SGNH hydrolase-like protein, which while structurally similar to the N-terminal domain of AlgX exhibits a distinctly different electrostatic surface potential. Consistent with other SGNH hydrolases, we identified a conserved catalytic triad composed of D190, H192 and S288 and demonstrated that AlgJ exhibits acetylesterase activity in vitro. Residues in the AlgJ signature motifs were found to form an extensive network of interactions that are critical for O-acetylation of alginate in vivo. Using two different electrospray ionization mass spectrometry (ESI-MS) assays we compared the abilities of AlgJ and AlgX to bind and acetylate alginate. Binding studies using defined length polymannuronic acid revealed that AlgJ exhibits either weak or no detectable polymer binding while AlgX binds polymannuronic acid specifically in a length-dependent manner. Additionally, AlgX was capable of utilizing the surrogate acetyl-donor 4-nitrophenyl acetate to catalyze the O-acetylation of polymannuronic acid. Our results, combined with previously published in vivo data, suggest that the annotated O-acetyltransferases AlgJ and AlgX have separate and distinct roles in O-acetylation. Our refined model for alginate acetylation places AlgX as the terminal acetlytransferase and provides a rationale for the variability in the number of proteins required for polysaccharide O-acetylation.

show abstract

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

P. aeruginosa SGNH Hydrolase-Like Proteins AlgJ and AlgX Have Similar Topology but Separate and Distinct Roles in Alginate Acetylation

et al. 2014

Self Cite

View full text Add to dashboard Cite

show abstract

“…Working on expanding this promising technology, Seeberger and co-workers approached the synthesis of oligosaccharides containing other important residues and linkages, including aminosugars [91][92][93][94], sialic acids [95], furanosides [96], 1,2-cis glycosides [97], glycopeptides [98], and branched oligosaccharide sequences [69]. Codee et al reported the synthesis of β-mannosides [99][100] using essentially the same automation platform. The acquired knowledge was applied to a very effective synthesis of Globo-H hexasaccharide.…”

Section: Peptide Synthesizer-based Automation Of Polymer-supported Symentioning

confidence: 99%

Automated Chemical Synthesis of Oligosaccharides and Glycoconjugates

Pistorio

Stine

Demchenko

2015

Carbohydrate Chemistry: State of the Art and Challenges for Drug Development

View full text Add to dashboard Cite

“…3,4 Ever more complex molecules are being assembled in an automated manner, and recent highlights include the assembly of libraries of plant-derived branched arabino-xylan and xyloglucan structures, 5 hyaluronic acid fragments up to 15 monosaccharides in length, 6 a 50-mer polymannoside, 7 a set of dermatan 8 and keratan sulfates, 9 a set of α-glucans, 10 and a collection of mannuronic acid alginates, built up to 12 β-mannuronic acid residues linkages. 11 These synthetic successes have shown that linear and branches structures can be assembled in an automated means and that both 1,2- trans and 1,2- cis linkages can be reliably installed using solid-phase chemistry. The method is especially attractive for the generation of libraries of oligosaccharides and oligosaccharides featuring repetitive elements.…”

Section: Introductionmentioning

confidence: 99%

Cyanopivaloyl Ester in the Automated Solid-Phase Synthesis of Oligorhamnans

et al. 2017

Self Cite

View full text Add to dashboard Cite

The development of effective protecting group chemistry is an important driving force behind the progress in the synthesis of complex oligosaccharides. Automated solid-phase synthesis is an attractive technique for the rapid assembly of oligosaccharides, built up of repetitive elements. The fact that (harsh) reagents are used in excess in multiple reaction cycles makes this technique extra demanding on the protecting groups used. Here, the synthesis of a set of oligorhamnan fragments is reported using the cyanopivaloyl (PivCN) ester to ensure effective neighboring group participation during the glycosylation events. The PivCN group combines the favorable characteristics of the parent pivaloyl (Piv) ester, stability, minimal migratory aptitude, minimal orthoester formation, while it can be cleaved under mild conditions. We show that the remote CN group in the PivCN renders the PivCN carbonyl more electropositive and thus susceptible to nucleophilic cleavage. This feature is built upon in the automated solid-phase assembly of the oligorhamnan fragments. Where the use of a Piv-protected building block failed because of incomplete cleavage, PivCN-protected synthons performed well and allowed the generation of oligorhamnans, up to 16 monosaccharides in length.

show abstract

Automated Solid‐Phase Synthesis of β‐Mannuronic Acid Alginates

Cited by 100 publications

References 30 publications

P. aeruginosa SGNH Hydrolase-Like Proteins AlgJ and AlgX Have Similar Topology but Separate and Distinct Roles in Alginate Acetylation

P. aeruginosa SGNH Hydrolase-Like Proteins AlgJ and AlgX Have Similar Topology but Separate and Distinct Roles in Alginate Acetylation

Automated Chemical Synthesis of Oligosaccharides and Glycoconjugates

Cyanopivaloyl Ester in the Automated Solid-Phase Synthesis of Oligorhamnans

Contact Info

Product

Resources

About