Structure of human ST8SiaIII sialyltransferase provides insight into cell-surface polysialylation

Volkers, Gesa; Worrall, L.J.; Kwan, David H.; Yu, Ching‐Ching; Baumann, Lars; Lameignère, Emilie; Wasney, Gregory A.; Scott, Nichollas E.; Wakarchuk, Warren W.; Foster, Leonard J.; Withers, Stephen G.; Strynadka, N.C.J.

doi:10.1038/nsmb.3060

Cited by 71 publications

(89 citation statements)

References 75 publications

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“…6 and Supplementary Table 5). This revealed a single Rossmann-like (GT-A variant 2) fold similar to other known GT29 sialyltransferases9,17,24,25, especially within the conserved “sialylmotif” sequence elements (Supplementary Fig. 6).…”

Section: Resultsmentioning

confidence: 62%

“…6). While structures have been reported for three of the four vertebrate GT29 sialyltransferase subfamilies that generate NeuAc-α2,6Gal17,24, NeuAc-α2,3Gal9, and NeuAc-α2,8NeuAc25 linkages, the present ST6GALNAC2 structure is the first reported for a NeuAc-α2,6GalNAc subfamily member17. Efforts to identify enzyme-CMP-acceptor structural co-complexes and perform kinetic analyses of site directed mutants are presently underway and will be reported elsewhere.…”

Section: Resultsmentioning

confidence: 84%

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Expression system for structural and functional studies of human glycosylation enzymes

et al. 2017

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Vertebrate glycoproteins and glycolipids are synthesized in complex biosynthetic pathways localized predominantly within membrane compartments of the secretory pathway. The enzymes that catalyze these reactions are exquisitely specific, yet few have been extensively characterized due to challenges associated with their recombinant expression as functional products. We used a modular approach to create an expression vector library encoding all known human glycosyltransferases, glycoside hydrolases, sulfotransferases, and other glycan modifying enzymes. We then expressed the enzymes as secreted catalytic domain fusion proteins in mammalian and insect cell hosts, purified and characterized a subset of the enzymes, and determined the structure of one, the sialyltransferase ST6GALNAC2. Many enzymes were produced at high yields and similar levels in both hosts, but individual protein expression levels varied widely. This expression vector library will be a transformative resource for recombinant enzyme production, broadly enabling structure-function studies and expanding applications of these enzymes in glycochemistry and glycobiology.

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Section: Resultsmentioning

confidence: 62%

Section: Resultsmentioning

confidence: 84%

Expression system for structural and functional studies of human glycosylation enzymes

et al. 2017

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“…Notably Mh PST shows no structural similarity to mammalian PSTs of the GT29 family, as the structure of the human ST8SiaIII enzyme exhibits a GT-A fold consisting of a single Rossmann-like domain 35 .…”

Section: Resultsmentioning

confidence: 99%

“…Recent structures of the human ST8SiaIII enzyme provided important insights into polysialylation in mammals 35 , but bacterial PSTs belong to a completely distinct family of glycosyltransferases (Carbohydrate Active Enzyme (CAZy) designated family GT38 distinct from GT29 for ST8SiaIII) 36 , which is not specific for particular acceptor proteins, but assembles polySia on a lipid-linked oligosaccharide 10, 37 .…”

Section: Introductionmentioning

confidence: 99%

X-ray crystallographic structure of a bacterial polysialyltransferase provides insight into the biosynthesis of capsular polysialic acid

Lizak

Worrall

Baumann

et al. 2017

Sci Rep

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Polysialic acid (polySia) is a homopolymeric saccharide that is associated with some neuroinvasive pathogens and is found on selective cell types in their eukaryotic host. The presence of a polySia capsule on these bacterial pathogens helps with resistance to phagocytosis, cationic microbial peptides and bactericidal antibody production. The biosynthesis of bacterial polySia is catalysed by a single polysialyltransferase (PST) transferring sialic acid from a nucleotide-activated donor to a lipid-linked acceptor oligosaccharide. Here we present the X-ray structure of the bacterial PST from Mannheimia haemolytica serotype A2, thereby defining the architecture of this class of enzymes representing the GT38 family. The structure reveals a prominent electropositive groove between the two Rossmann-like domains forming the GT-B fold that is suitable for binding of polySia chain products. Complex structures of PST with a sugar donor analogue and an acceptor mimetic combined with kinetic studies of PST active site mutants provide insight into the principles of substrate binding and catalysis. Our results are the basis for a molecular understanding of polySia biosynthesis in bacteria and might assist the production of polysialylated therapeutic reagents and the development of novel antibiotics.

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“…The crystal structure of ST8SiaIII, which is discussed to build sialic acid oligomers [4], support the models of Frederic Troy and Karen Colley [52]. Strynadka and colleagues compared—on the basis of their crystal structure—the amino acid sequences of ST8SiaII and IV with ST8SiaIII.…”

Section: Polysialylation Of Ncammentioning

confidence: 93%

Is Polysialylated NCAM Not Only a Regulator during Brain Development But also during the Formation of Other Organs?

Galuska

Lütteke²,

Galuska

2017

Biology

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In mammals several cell adhesion molecules are involved during the pre- and postnatal development of all organ systems. A very prominent member of this family is the neural cell adhesion molecule (NCAM). Interestingly, NCAM can be a target for a special form of posttranslational modification: polysialylation. Whereas nearly all extracellular proteins bear mono-sialic acid residues, only a very small group can be polysialylated. Polysialic acid is a highly negatively-charged sugar polymer and can comprise more than 90 sialic acid residues in postnatal mouse brains increasing dramatically the hydrodynamic radius of their carriers. Thus, adhesion and communication processes on cell surfaces are strongly influenced allowing, e.g., the migration of neuronal progenitor cells. In the developing brain the essential role of polysialylated NCAM has been demonstrated in many studies. In comparison to the neuronal system, however, during the formation of other organs the impact of the polysialylated form of NCAM is not well characterized and the number of studies is limited so far. This review summarizes these observations and discusses possible roles of polysialylated NCAM during the development of organs other than the brain.

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Structure of human ST8SiaIII sialyltransferase provides insight into cell-surface polysialylation

Cited by 71 publications

References 75 publications

Expression system for structural and functional studies of human glycosylation enzymes

Expression system for structural and functional studies of human glycosylation enzymes

X-ray crystallographic structure of a bacterial polysialyltransferase provides insight into the biosynthesis of capsular polysialic acid

Is Polysialylated NCAM Not Only a Regulator during Brain Development But also during the Formation of Other Organs?

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