Structure and mechanism of TagA, a novel membrane-associated glycosyltransferase that produces wall teichoic acids in pathogenic bacteria

Kattke, Michele D.; Gosschalk, Jason E.; Martinez, Orlando E.; Kumar, Garima; Gale, Robert T.; Cascio, Duilio; Sawaya, M.R.; Philips, Martin; Brown, Eric D.; Clubb, Robert T.

doi:10.1371/journal.ppat.1007723

Cited by 25 publications

(46 citation statements)

References 53 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…The Leloir glycosyltransferases containing a GT-D fold catalyze the transfer of glucose to hexasaccharide O -linked to serine-rich repeats of bacterial adhesins [39]. The most recent addition, is the N -acetyl- d -mannose transferase utilizing non-Leloir undecaprenyl-linked glycosyl diphosphates with a unique GT-E fold [40].…”

Section: Glycosyltransferases In Naturementioning

confidence: 99%

Leloir Glycosyltransferases in Applied Biocatalysis: A Multidisciplinary Approach

Mestrom

Przypis

Kowalczykiewicz

et al. 2019

IJMS

View full text Add to dashboard Cite

Enzymes are nature's catalyst of choice for the highly selective and efficient coupling of carbohydrates. Enzymatic sugar coupling is a competitive technology for industrial glycosylation reactions, since chemical synthetic routes require extensive use of laborious protection group manipulations and often lack regio-and stereoselectivity. The application of Leloir glycosyltransferases has received considerable attention in recent years and offers excellent control over the reactivity and selectivity of glycosylation reactions with unprotected carbohydrates, paving the way for previously inaccessible synthetic routes. The development of nucleotide recycling cascades has allowed for the efficient production and reuse of nucleotide sugar donors in robust one-pot multi-enzyme glycosylation cascades. In this way, large glycans and glycoconjugates with complex stereochemistry can be constructed. With recent advances, LeLoir glycosyltransferases are close to being applied industrially in multi-enzyme, programmable cascade glycosylations.hydroxynitrile lyases catalyzed the enzymatic hydrolysis of the glycoside amygdalin [3]. Moving almost two centuries forward, the largest volumetric biocatalytic industrial process is the application of glucose isomerase for the production of high fructose syrup for food and drink applications, producing fructose from glucose at 10 7 tons per year [4]. The secret of the success of enzymes in the production or treatment of carbohydrates and glycosides is their exquisite stereo-and regioselectivity. The excellent selectivity of enzymes is required due to the diversity of structural features of carbohydrates [5], comprising d-and l-epimers, ring size, anomeric configuration, linkages, branching, and oxidation state(s). Since drug targets often exhibit specificity for all of these structural features, the production process should not contain any side-products to prevent undesired side-effects [6].The challenge in the synthesis of carbohydrates is their wide variety of functionalities and stereochemistry ( Figure 1). (Poly)hydroxyaldehydes containing a terminal aldehyde are referred to as aldoses and (poly)hydroxyketones are defined as ketoses. In aqueous solutions, monosaccharides form equilibrium mixtures of linear open-chain and ring-closed 5-or 6-membered furanoses or pyranoses, respectively. For aldoses, the asymmetric ring forms at C-1. For ketoses, it closes at C-2 as an axial (α) or equatorial (β) hemiacetal or hemiketal, respectively (commonly defined as the anomeric center). A glycosidic linkage is a covalent O-, S-, N-, or C-bond connecting a monosaccharide to another residue resulting in a glycoside, while glucoside is specific for a glucose moiety. The equatorial or axial position of the glycosidic bond is referred to as α-(axial) or β-linkage (equatorial). The number of carbohydrates linked via glycosidic bonds can be subdivided into oligosaccharides with two to ten linked carbohydrates, while polysaccharides (glycans) contain more than ten glycosidic bonds. A glycan either con...

show abstract

Section: Glycosyltransferases In Naturementioning

confidence: 99%

Leloir Glycosyltransferases in Applied Biocatalysis: A Multidisciplinary Approach

Mestrom

Przypis

Kowalczykiewicz

et al. 2019

IJMS

View full text Add to dashboard Cite

show abstract

“…Most of the GTs whose 3D structure has been determined experimentally adopt either the GT-A or GT-B fold; few others have GT-C, GT-D or GT-E fold (42–44). Such a conservation of fold despite of sequence and functional differences is not surprising since it has long been known that 3D structures are more conserved than sequences.…”

Section: Resultsmentioning

confidence: 99%

Searching sequence databases for functional homologs using profile HMMs: how to set bit score thresholds?

Srivastava

Hembrom

Kumawat

et al. 2021

Preprint

View full text Add to dashboard Cite

UniProt and BFD databases together have 2.5 billion protein sequences. A large majority of these proteins have been electronically annotated. Automated annotation pipelines, vis-à-vis manual curation, have the advantage of scale and speed but are fraught with relatively higher error rates. This is because sequence homology does not necessarily translate to functional homology, molecular function specification is hierarchic and not all functional families have the same amount of experimental data that one can exploit for annotation. Consequently, customization of annotation workflow is inevitable to minimize annotation errors. In this study, we illustrate possible ways of customizing the search of sequence databases for functional homologs using profile HMMs. Choosing an optimal bit score threshold is a critical step in the application of HMMs. We illustrate ways in which an optimal bit score can be arrived at using four Case Studies. These are the single domain nucleotide sugar 6-dehydrogenase and lysozyme-C families, and SH3 and GT-A domains which are typically found as a part of multi-domain proteins. We also discuss the limitations of using profile HMMs for functional annotation and suggests some possible ways to partially overcome such limitations.

show abstract

“…The dual-activity mannosyltransferase/ phosphorylases of family GT108 have the highest mRE (0.281) and have indeed been shown to adopt a unique five-bladed β-propeller fold that is completely different from the four GT folds 40 . Another family predicted to have a novel fold, GT26, has a single representative crystal structure for a membrane associated GT TagA, from a bacteria T. italicus , which also adopts a novel fold 41 . Here, we predict three additional families, the fungal β-1,2-mannosyltransferases Bmt/Wry (GT91), plant peptidyl serine α-galactosyltransferases Sgt (GT96) and bacterial α-2,6-sialyltransferases (GT97), that likely adopt novel GT folds as well.…”

Section: Resultsmentioning

confidence: 99%

Mapping the glycosyltransferase fold landscape using deep learning

Taujale¹,

Zhou²,

Yeung³

et al. 2021

Preprint

View full text Add to dashboard Cite

Glycosyltransferases (GTs) play fundamental roles in nearly all cellular processes through 10 the biosynthesis of complex carbohydrates and glycosylation of diverse protein and small 11 molecule substrates. The extensive structural and functional diversification of GTs presents a 12 major challenge in mapping the relationships connecting sequence, structure, fold and function 13 using traditional bioinformatics approaches. Here, we present a convolutional neural network 14 with attention (CNN-attention) based deep learning model that leverages simple secondary 15 structure representations generated from primary sequences to provide GT fold prediction with 16 high accuracy. The model learned distinguishing features free of primary sequence alignment 17 constraints and, unlike other models, is highly interpretable and helped identify common 18 secondary structural features shared by divergent families. The model delineated sequence and 19 structural features characteristic of individual fold types, while classifying them into distinct 20 clusters that group evolutionarily divergent families based on shared secondary structural 21 features. We further extend our model to classify GT families of unknown folds and variants of 22 known folds. By identifying families that are likely to adopt novel folds such as GT91, GT96 and 23 GT97, our studies identify targets for future structural studies and expand the GT fold landscape.

show abstract

Structure and mechanism of TagA, a novel membrane-associated glycosyltransferase that produces wall teichoic acids in pathogenic bacteria

Cited by 25 publications

References 53 publications

Leloir Glycosyltransferases in Applied Biocatalysis: A Multidisciplinary Approach

Leloir Glycosyltransferases in Applied Biocatalysis: A Multidisciplinary Approach

Searching sequence databases for functional homologs using profile HMMs: how to set bit score thresholds?

Mapping the glycosyltransferase fold landscape using deep learning

Contact Info

Product

Resources

About