Structural insight into mammalian sialyltransferases

Rao, Francesco; Rich, Justin; Rakić, Bojana; Buddai, Sai K.; Schwartz, Marc F.; Johnson, Karl; Bowe, Caryn; Wakarchuk, Warren W.; DeFrees, Shawn; Withers, Stephen G.; Strynadka, N.C.J.

doi:10.1038/nsmb.1685

Cited by 107 publications

(148 citation statements)

References 22 publications

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“…The equivalent disordered segment in monomer B is larger, comprising 12 residues (residues 353-366) as well as an adjacent disordered segment of residues 346 -347. These regions resemble the disordered "lid" regions capping the sugarnucleotide donor-binding site on other glycosyltransferases (67,68), including ST3GAL1 (25), that typically become structured upon binding the sugar-nucleotide donor.…”

Section: Resultsmentioning

confidence: 99%

“…A varied "stem" domain acts as a linker between the membrane anchor and the more conserved catalytic domain. Because all mammalian sialyltransferases can employ CMP-Neu5Ac as a sugar donor, the conserved sialylmotif sequences were proposed to include the donor binding site, and this was subsequently confirmed by mutagenesis studies (24) and the structure of ST3GAL1 containing bound CMP (25). In contrast, the broad differences in acceptor specificity across the sialyltransferase family were reflected in divergent sequences outside the sialylmotifs (20,22).…”

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confidence: 91%

“…In contrast, bacterial sialyltransferases are found in distinct CAZy families (GT30, GT38, GT42, GT73, and GT80) where they transfer mono-or polysialic acid or related KDO residues (23). Minimal sequence identity has been observed between animal and bacterial sialyltransferases, but structural similarity was observed between some bacterial GT42 sialyltransferases and the mammalian sialyltransferases within the sialylmotif region (25).…”

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confidence: 93%

“…The structure revealed several conserved features with ST3GAL1 (25), the bacterial GT42 sialyltransferase, CstII (30), and the recently published structure of human ST6GAL1 (29), including the sialylmotif region involved in binding the sugar-nucleotide donor. Modeling, molecular dynamics (MD) 5 simulations, and kinetic analysis of site-directed mutants provide evidence for a broadly similar positioning of sugar donor, glycan acceptor, and catalytic base for all four enzymes.…”

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confidence: 99%

“…Insights into the domain structures and specificities of the mammalian sialyltransferases have come from extensive enzymatic studies on wild type and mutant enzymes (20,26,27), from the crystal structures of a pig ST3GAL1 involved in the synthesis of Neu5Ac␣2,3Gal linkages on O-linked glycans (25), and a structure of human ST6GAL1 that was published while this manuscript was being completed (29). The overall domain organization among the sialyltransferases is similar (28).…”

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confidence: 99%

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Enzymatic Basis for N-Glycan Sialylation

Forouhar

Thieker

et al. 2013

Journal of Biological Chemistry

118

130

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Background: Specificity and enzymology of glycan sialylation is poorly understood, despite its importance in biological recognition. Results: ST6GAL1 structure was determined, and substrate binding was modeled to probe active site specificity. Conclusion:The structure provides insights into the enzymatic basis of glycan sialylation. Significance: Knowledge of the enzyme structure can lead to broader understanding of enzymatic sialylation and selective inhibitor design.

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Section: Resultsmentioning

confidence: 99%

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confidence: 91%

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confidence: 93%

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confidence: 99%

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confidence: 99%

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Enzymatic Basis for N-Glycan Sialylation

Forouhar

Thieker

et al. 2013

Journal of Biological Chemistry

118

130

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Engineering of a Cytidine 5′‐Monophosphate‐Sialic Acid Synthetase for Improved Tolerance to Functional Sialic Acids

Dong

Kickstein

et al. 2013

Adv Synth Catal

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Sialic acid-containing glycoconjugates at the cell surface are of high importance in carbohydrate-mediated recognition phenomena in physiological and pathological events, as well as in bacterial or viral infection. A key step in the enzymatic synthesis of natural sialoconjugates and functional synthetic analogues is the activation of sialic acids to cytidine 5'-monophosphate (CMP)-sialic acid intermediates catalyzed by CMP-sialic acid synthetase (CSS). Based on our recently developed aligned protein model of substrate binding and a simple colorimetric screening assay, we have engineered the CSS from Neisseria meningitidis by structure-guided site-specific saturation mutagenesis at positions 192/193 to generate enzymes with broadened substrate scope. Top hits, including the F192S/F193Y variant, display an improvement of up to 70-fold catalytic efficiency relative to wild-type CSS for the conversion of sterically demanding N-acyl modified sialic acid analogues, without compromising protein stability. Such significantly enhanced substrate capacity is a major step forward to realizing a generalized chemo-enzymatic strategy for the efficient preparation of neo-sialoconjugate libraries, demonstrated by the highly efficient, regio-and stereospecific synthesis of 2,6-sialyllactose analogues by enzymatic coupling to the highly substrate tolerant a2,6-sialyltransferase from Photobacterium leiognathi JT-SHIZ-145. Our results further document the unusual versatility of the N. meningitidis CSS and engineered variants for a common synthetic approach to sialoconjugates comprising a large diversity of natural and non-natural sialic acid forms without the need for post-synthetic enzymatic modification.

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Recent Developments in Enzymatic Synthesis of Modified Sialic Acid Derivatives

Withers

2015

Adv Synth Catal

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Sialic acid-containing glycans play important roles in biology, but their synthesisb ys tandard chemical approaches is challenging. Enzymatic assemblyi sg enerally ab etter approach. In the last two decades,anumber of bacterial sialyltransferases (STs) and trans-sialidasesh ave been identifieda nd an umber of them found to exhibit broad substrate specificity.I nt his review,w ew illf ocuso nt he donor and acceptor substrate specificities of these enzymes along with the enzymatic routes employed to prepare sialosidesb earing modificationsa ts pecific positions on the sialic acid carbon skeleton. Understanding the substrate tolerance of these enzymes will help researchers to choose the best biocatalyst for the assemblyo fs pecific sialosides,w hich can be valuable tools in the study of or inhibition of sialidases and STs and sialica cid-binding proteins.I na ddition,t he ability to study these processes using synthetic sialylated glycans will leadt oagreater understanding of their biologya nd willl ead to newt herapeutic targets.1I ntroduction 2S ialic AcidsinN ature 3S ynthesis of Sialic Acid-ContainingO ligosaccharides 4B acterial Sialyltransferases and trans-Sialidases 4

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Structural insight into mammalian sialyltransferases

Cited by 107 publications

References 22 publications

Enzymatic Basis for N-Glycan Sialylation

Enzymatic Basis for N-Glycan Sialylation

Engineering of a Cytidine 5′‐Monophosphate‐Sialic Acid Synthetase for Improved Tolerance to Functional Sialic Acids

Recent Developments in Enzymatic Synthesis of Modified Sialic Acid Derivatives

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