Structures of Complexes of a Metal-independent Glycosyltransferase GT6 from Bacteroides ovatus with UDP-N-Acetylgalactosamine (UDP-GalNAc) and Its Hydrolysis Products

Pham, Thuy Thi Thu; Stinson, Brittany; Thiyagarajan, N.; Lizotte-Waniewski, Michelle; Brew, Keith; Acharya, K. Ravi

doi:10.1074/jbc.m113.545384

Cited by 17 publications

(8 citation statements)

References 33 publications

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“…3 ) ( 7 , 12 , 15 – 18 , 20 , 22 – 24 ). Additionally, this GalNAc conformation in AAGlyB is also different from the GalNAc conformation recently described in the similar, but metal-independent GT (CAZy group GT6) from Bacteroides ovatus ( 21 ).…”

Section: Resultscontrasting

confidence: 80%

“…Importantly, the structure of the AAGlyB- 3 complex contains the donor analogue with a partially intact GalNAc portion, which, for the first time, gives an indication of the binding mode of UDP-GalNAc to a human blood group GT. In this new structure, the GalNAc moiety is placed in the active site in a conformation, which is different from the canonical tucked under conformation previously observed in other UDP-sugar-GT complex structures ( 7 , 12 , 15 – 18 , 20 – 24 ).…”

Section: Introductionmentioning

confidence: 58%

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Novel UDP-GalNAc Derivative Structures Provide Insight into the Donor Specificity of Human Blood Group Glycosyltransferase

Wagner

Pesnot

Palcic

et al. 2015

Journal of Biological Chemistry

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Two closely related glycosyltransferases are responsible for the final step of the biosynthesis of ABO(H) human blood group A and B antigens. The two enzymes differ by only four amino acid residues, which determine whether the enzymes transfer GalNAc from UDP-GalNAc or Gal from UDP-Gal to the H-antigen acceptor. The enzymes belong to the class of GT-A folded enzymes, grouped as GT6 in the CAZy database, and are characterized by a single domain with a metal dependent retaining reaction mechanism. However, the exact role of the four amino acid residues in the specificity of the enzymes is still unresolved. In this study, we report the first structural information of a dual specificity cis-AB blood group glycosyltransferase in complex with a synthetic UDP-GalNAc derivative. Interestingly, the GalNAc moiety adopts an unusual yet catalytically productive conformation in the binding pocket, which is different from the “tucked under” conformation previously observed for the UDP-Gal donor. In addition, we show that this UDP-GalNAc derivative in complex with the H-antigen acceptor provokes the same unusual binding pocket closure as seen for the corresponding UDP-Gal derivative. Despite this, the two derivatives show vastly different kinetic properties. Our results provide a important structural insight into the donor substrate specificity and utilization in blood group biosynthesis, which can very likely be exploited for the development of new glycosyltransferase inhibitors and probes.

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Section: Resultscontrasting

confidence: 80%

Section: Introductionmentioning

confidence: 58%

Novel UDP-GalNAc Derivative Structures Provide Insight into the Donor Specificity of Human Blood Group Glycosyltransferase

Wagner

Pesnot

Palcic

et al. 2015

Journal of Biological Chemistry

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“…The remaining six strains have 1–11 of these proteins. These glycosyltransferases function in the biosynthesis of polysaccharides and complex oligosaccharides and may play roles in various biological functions such as cell signaling, cellular interactions and pathogenesis [27].…”

Section: Resultsmentioning

confidence: 99%

Comparative genomics of transport proteins in seven Bacteroides species

Zafar

Saier

2018

PLoS ONE

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The communities of beneficial bacteria that live in our intestines, the gut microbiome, are important for the development and function of the immune system. Bacteroides species make up a significant fraction of the human gut microbiome, and can be probiotic and pathogenic, depending upon various genetic and environmental factors. These can cause disease conditions such as intra-abdominal sepsis, appendicitis, bacteremia, endocarditis, pericarditis, skin infections, brain abscesses and meningitis. In this study, we identify the transport systems and predict their substrates within seven Bacteroides species, all shown to be probiotic; however, four of them (B. thetaiotaomicron, B. vulgatus, B. ovatus, B. fragilis) can be pathogenic (probiotic and pathogenic; PAP), while B. cellulosilyticus, B. salanitronis and B. dorei are believed to play only probiotic roles (only probiotic; OP). The transport system characteristics of the four PAP and three OP strains were identified and tabulated, and results were compared among the seven strains, and with E. coli and Salmonella strains. The Bacteroides strains studied contain similarities and differences in the numbers and types of transport proteins tabulated, but both OP and PAP strains contain similar outer membrane carbohydrate receptors, pore-forming toxins and protein secretion systems, the similarities were noteworthy, but these Bacteroides strains showed striking differences with probiotic and pathogenic enteric bacteria, particularly with respect to their high affinity outer membrane receptors and auxiliary proteins involved in complex carbohydrate utilization. The results reveal striking similarities between the PAP and OP species of Bacteroides, and suggest that OP species may possess currently unrecognized pathogenic potential.

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“…Mammalian members of family GT-6 are metal dependent, and contain the DXD motif, whereas a number of the bacterial species assigned to the same family have a conserved NXN motif and are metal independent. The structural basis for this has been described with two family GT-6 structures from Bacteroides ovatus [ 51,52 ]. There is a high degree of structural homology between the bacterial and mammalian GT-6 enzymes, with equivalent residues for binding substrates.…”

Section: Other Highlightsmentioning

confidence: 99%