Two complementary α-fucosidases from Streptococcus pneumoniae promote complete degradation of host-derived carbohydrate antigens

Hobbs, Joanne K.; Pluvinage, B.; Robb, Melissa; Smith, Steven P.; Boraston, A.B.

doi:10.1074/jbc.ra119.009368

Cited by 20 publications

(25 citation statements)

References 70 publications

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“…However, its presence does not always show as utilisation of 2 0 -FL (42) . Recently, Streptococcus pneumoniae has been shown to contain two α-fucosidases (43) . The genome sequence of S. mutans strain UA159 is known to have four ABC transporters (44) , but transcription profiles for fucose or HMOs were not studied.…”

Section: Discussionmentioning

confidence: 99%

Influence of 2′-fucosyllactose and galacto-oligosaccharides on the growth and adhesion of Streptococcus mutans

et al. 2020

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Abstract Human milk oligosaccharides, such as 2′-fucosyllactose (2′-FL), and galacto-oligosaccharides (GOS), a prebiotic carbohydrate mixture, are being increasingly added to infant formulas, necessitating the understanding of their impact on the oral microbiota. Here, for the first time, the effects of 2′-FL and GOS on the planktonic growth and adhesion characteristics of the caries-associated oral pathogen Streptococcus mutans were assessed, and the results were compared against the effects of xylitol, lactose and glucose. There were differences in S. mutans growth between 2′-FL and GOS. None of the three S. mutans strains grew with 2′-FL, while they all grew with GOS as well as lactose and glucose. Xylitol inhibited S. mutans growth. The adhesion of S. mutans CI 2366 to saliva-coated hydroxyapatite was reduced by 2′-FL and GOS. Exopolysaccharide-mediated adhesion of S. mutans DSM 20523 to a glass surface was decreased with 2′-FL, GOS and lactose, and the adhesion of strain CI 2366 strain was reduced only by GOS. Unlike GOS, 2′-FL did not support the growth of any S. mutans strain. Neither 2′-FL nor GOS enhanced the adhesive properties of the S. mutans strains, but they inhibited some of the tested strains. Thus, the cariogenic tendency may vary between infant formulas containing different types of oligosaccharides.

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Section: Discussionmentioning

confidence: 99%

Influence of 2′-fucosyllactose and galacto-oligosaccharides on the growth and adhesion of Streptococcus mutans

et al. 2020

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“…org). The macromolecular architecture is conserved with [22] and Bifidobacterium longum subsp. infantis (RMSD: 1.128 Å) [67].…”

Section: R Gnavus Gh29 E1_10125 Fucosidase Can Accommodate Terminal mentioning

confidence: 99%

Fucosidases from the human gut symbiont Ruminococcus gnavus

Rebello

Crost

et al. 2020

Cell. Mol. Life Sci.

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The availability and repartition of fucosylated glycans within the gastrointestinal tract contributes to the adaptation of gut bacteria species to ecological niches. To access this source of nutrients, gut bacteria encode α-l-fucosidases (fucosidases) which catalyze the hydrolysis of terminal α-l-fucosidic linkages. We determined the substrate and linkage specificities of fucosidases from the human gut symbiont Ruminococcus gnavus. Sequence similarity network identified strain-specific fucosidases in R. gnavus ATCC 29149 and E1 strains that were further validated enzymatically against a range of defined oligosaccharides and glycoconjugates. Using a combination of glycan microarrays, mass spectrometry, isothermal titration calorimetry, crystallographic and saturation transfer difference NMR approaches, we identified a fucosidase with the capacity to recognize sialic acid-terminated fucosylated glycans (sialyl Lewis X/A epitopes) and hydrolyze α1-3/4 fucosyl linkages in these substrates without the need to remove sialic acid. Molecular dynamics simulation and docking showed that 3′-Sialyl Lewis X (sLeX) could be accommodated within the binding site of the enzyme. This specificity may contribute to the adaptation of R. gnavus strains to the infant and adult gut and has potential applications in diagnostic glycomic assays for diabetes and certain cancers.

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“…ride). Prior defucosylation of this tetrasaccharide into the linear form would allow Aga to act; however, the two characterized pneumococcal ␣-fucosidases, which are the only known fucosidases in this bacterium, lack this activity (44). Furthermore, we found Aga to have an intracellular location, which makes a potential role in host glycan processing even less likely.…”

Section: Molecular Analysis Of Pneumococcal Rfo Utilizationmentioning

confidence: 74%

“…Coordination is similar between both residues, with interactions between Trp 274 and O5-C1-C2 plane of galactose/glucose. Direct hydrogen bonds are made between O4, O3, O2, and Asp 308 , Lys 42 , and Glu 44 , respectively.…”

Section: Molecular Analysis Of Pneumococcal Rfo Utilizationmentioning

confidence: 99%

“…Protein expression constructs were transformed into BL21(DE3). Expression of Aga, RafE, and GtfA was performed in LB broth with 0.5 mM isopropyl ␤-D-1-thiogalactopyranoside induction at 16°C for 18 h. Standard procedures, as previously detailed (44), were used to lyse cells and purify the released proteins by immobilized metal affinity chromatography. Subsequent purification by size-exclusion chromatography, using either an S100 or S200 HiPrep 16/60 Sephacryl column (GE Healthcare) as appropriate, was performed using 20 mM Tris, pH 8.0, 500 mM NaCl.…”

Section: Protein Expression and Purificationmentioning

confidence: 99%

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Molecular analysis of an enigmatic Streptococcus pneumoniae virulence factor: The raffinose-family oligosaccharide utilization system

Hobbs¹,

Meier²,

Pluvinage³

et al. 2019

Journal of Biological Chemistry

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Streptococcus pneumoniae is an opportunistic respiratory pathogen that can spread to other body sites, including the ears, brain, and blood. The ability of this bacterium to break down, import, and metabolize a wide range of glycans is key to its virulence. Intriguingly, S. pneumoniae can utilize several plant oligosaccharides for growth in vitro, including raffinose-family oligosaccharides (RFOs, which are α-(1→6)-galactosyl extensions of sucrose). An RFO utilization locus has been identified in the pneumococcal genome; however, none of the proteins encoded by this locus have been biochemically characterized. The enigmatic ability of S. pneumoniae to utilize RFOs has recently received attention because mutations in two of the RFO locus genes have been linked to the tissue tropism of clinical pneumococcal isolates. Here, we use functional studies combined with X-ray crystallography to show that although the pneumococcal RFO locus encodes for all the machinery required for uptake and degradation of RFOs, the individual pathway components are biochemically inefficient. We also demonstrate that the initiating enzyme in this pathway, the α-galactosidase Aga (a family 36 glycoside hydrolase), can cleave α-(1→3)-linked galactose units from a linear blood group antigen. We propose that the pneumococcal RFO pathway is an evolutionary relic that is not utilized in this streptococcal species and, as such, is under no selection pressure to maintain binding affinity and/or catalytic efficiency. We speculate that the apparent contribution of RFO utilization to pneumococcal tissue tropism may, in fact, be due to the essential role the ATPase RafK plays in the transport of other carbohydrates.

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Two complementary α-fucosidases from Streptococcus pneumoniae promote complete degradation of host-derived carbohydrate antigens

Cited by 20 publications

References 70 publications

Influence of 2′-fucosyllactose and galacto-oligosaccharides on the growth and adhesion of Streptococcus mutans

Influence of 2′-fucosyllactose and galacto-oligosaccharides on the growth and adhesion of Streptococcus mutans

Fucosidases from the human gut symbiont Ruminococcus gnavus

Molecular analysis of an enigmatic Streptococcus pneumoniae virulence factor: The raffinose-family oligosaccharide utilization system

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