Quantifying the binding stoichiometry and affinity of histo-blood group antigen oligosaccharides for human noroviruses

AbstractHuman noroviruses (HuNoVs) are a major cause of acute gastroenteritis. Many HuNoVs recognize histo-blood group antigens (HBGAs) as cellular receptors or attachment factors for infection. It was recently proposed that HuNoV recognition of HBGAs involves a cooperative, multistep binding mechanism that exploits both known and previously unknown glycan binding sites. In this study, binding measurements, implemented using electrospray ionization mass spectrometry (ESI-MS) were performed on homodimers of the protruding domain (P dimers) of the capsid protein of three HuNoV strains [Saga (GII.4), Vietnam 026 (GII.10) and VA387 (GII.4)] with the ethyl glycoside of the B trisaccharide (α-d-Gal-(1→3)-[α-l-Fuc-(1→2)]-β-d-Gal-OC2H5) and free B type 1 tetrasaccharide (α-d-Gal-(1→3)-[α-l-Fuc-(1→2)]-β-d-Gal-(1→3)-d-GlcNAc) in an effort to confirm the existence of new HBGA binding sites. After correcting the mass spectra for nonspecific interactions that form in ESI droplets as they evaporate to dryness, all three P dimers were found to bind a maximum of two B trisaccharides at the highest concentrations investigated. The apparent affinities measured for stepwise binding of B trisaccharide suggest positive cooperativity. Similar results were obtained for B type 1 tetrasaccharide binding to Saga P dimer. Based on these results, it is proposed that HuNoV P dimers possess only two HBGA binding sites. It is also shown that nonspecific binding corrections applied to mass spectra acquired using energetic ion source conditions that promote in-source dissociation can lead to apparent HuNoV–HBGA oligosaccharide binding stoichiometries and affinities that are artificially high. Finally, evidence that high concentrations of oligosaccharide can induce conformational changes in HuNoV P dimers is presented.

Download Full-text

Investigation of Atmospheric Pressure Photochemical Ionization Mass Spectra of Binary Organic Solutions without Their Separation in Dependence on the Concentration of Solutions and Analyte Vapors in Nitrogen Using the Exponential Dilution Method and a Time-of-Flight Mass Spectrometer with an Atmospheric Pressure Photochemical Ionization Ion Source

Journal of Analytical Chemistry ◽

10.1134/s1061934820140117 ◽

2020 ◽

Vol 75 (14) ◽

pp. 1743-1748

Author(s):

I. A. Revelsky ◽

A. A. Sysoev ◽

A. I. Revelsky ◽

A. G. Adamov ◽

S. S. Poteshin ◽

...

Keyword(s):

Mass Spectrometer ◽

Atmospheric Pressure ◽

Mass Spectra ◽

Time Of Flight ◽

Ion Source ◽

Dilution Method ◽

Ionization Mass ◽

Concentration Of Solutions ◽

Exponential Dilution ◽

Organic Solutions

Download Full-text

Formation and destruction of the guanine quartet in solution observed by cold-spray ionization mass spectrometryElectronic supplementary information available: CSI and ESI mass spectra of dG, dC, dA and dT, and schematic diagram of the coldspray ion source. See http://www.rsc.org/suppdata/cc/b2/b212432g/

Chemical Communications ◽

10.1039/b212432g ◽

2003 ◽

pp. 788-789 ◽

Cited By ~ 24

Author(s):

Shigeru Sakamoto ◽

Kazuhiko Nakatani ◽

Isao Saito ◽

Kentaro Yamaguchi

Keyword(s):

Cold Spray ◽

Mass Spectra ◽

Ion Source ◽

Supplementary Information ◽

Ionization Mass ◽

Schematic Diagram ◽

Guanine Quartet

Download Full-text

Human Noroviruses' Fondness for Histo-Blood Group Antigens

Journal of Virology ◽

10.1128/jvi.02968-14 ◽

2014 ◽

Vol 89 (4) ◽

pp. 2024-2040 ◽

Cited By ~ 73

Author(s):

Bishal K. Singh ◽

Mila M. Leuthold ◽

Grant S. Hansman

Keyword(s):

Crystal Structures ◽

Blood Group ◽

Enteric Bacteria ◽

Blood Group Antigens ◽

Binding Interactions ◽

Human Noroviruses ◽

X Ray ◽

P Domain ◽

Lewis Y ◽

Binding Mechanisms

ABSTRACTHuman noroviruses are the dominant cause of outbreaks of gastroenteritis around the world. Human noroviruses interact with the polymorphic human histo-blood group antigens (HBGAs), and this interaction is thought to be important for infection. Indeed, synthetic HBGAs or HBGA-expressing enteric bacteria were shown to enhance norovirus infection in B cells. A number of studies have found a possible relationship between HBGA type and norovirus susceptibility. The genogroup II, genotype 4 (GII.4) noroviruses are the dominant cluster, evolve every other year, and are thought to modify their binding interactions with different HBGA types. Here we show high-resolution X-ray crystal structures of the capsid protruding (P) domains from epidemic GII.4 variants from 2004, 2006, and 2012, cocrystallized with a panel of HBGA types (H type 2, Lewis Y, Lewis B, Lewis A, Lewis X, A type, and B type). Many of the HBGA binding interactions were found to be complex, involving capsid loop movements, alternative HBGA conformations, and HBGA rotations. We showed that a loop (residues 391 to 395) was elegantly repositioned to allow for Lewis Y binding. This loop was also slightly shifted to provide direct hydrogen- and water-mediated bonds with Lewis B. We considered that the flexible loop modulated Lewis HBGA binding. The GII.4 noroviruses have dominated outbreaks over the past decade, which may be explained by their exquisite HBGA binding mechanisms, their fondness for Lewis HBGAs, and their temporal amino acid modifications.IMPORTANCEOur data provide a comprehensive picture of GII.4 P domain and HBGA binding interactions. The exceptionally high resolutions of our X-ray crystal structures allowed us to accurately recognize novel GII.4 P domain interactions with numerous HBGA types. We showed that the GII.4 P domain-HBGA interactions involved complex binding mechanisms that were not previously observed in norovirus structural studies. Many of the GII.4 P domain-HBGA interactions we identified were negative in earlier enzyme-linked immunosorbent assay (ELISA)-based studies. Altogether, our data show that the GII.4 norovirus P domains can accommodate numerous HBGA types.

Download Full-text

Association of Type 1 Blood Group Antigens with Urinary Tract Infections in Children with Genitourinary Structural Abnormalities

The Journal of Urology ◽

10.1016/s0022-5347(17)39492-2 ◽

1990 ◽

Vol 144 (2 Part 2) ◽

pp. 469-473 ◽

Cited By ~ 4

Author(s):

Joel Sheinfeld ◽

Carlos Cordon-Cardo ◽

William R. Fair ◽

David D. Wartinger ◽

Ronald Rabinowitz

Keyword(s):

Urinary Tract ◽

Blood Group ◽

Urinary Tract Infections ◽

Blood Group Antigens ◽

Structural Abnormalities ◽

Tract Infections

Download Full-text

Structural Constraints on Human Norovirus Binding to Histo-Blood Group Antigens

mSphere ◽

10.1128/msphere.00049-16 ◽

2016 ◽

Vol 1 (2) ◽

Cited By ~ 22

Author(s):

Bishal K. Singh ◽

Mila M. Leuthold ◽

Grant S. Hansman

Keyword(s):

Aspartic Acid ◽

Blood Group ◽

Blood Group Antigens ◽

Structural Constraints ◽

Human Norovirus ◽

Genotype 4 ◽

Human Noroviruses ◽

X Ray ◽

X Ray Crystallography ◽

New Findings

ABSTRACT Human norovirus interacts with the polymorphic human histo-blood group antigens (HBGAs), and this interaction is thought to be important for infection. The genogroup II genotype 4 (GII.4) noroviruses are the dominant cluster, evolve every other year, and are thought to modify their binding interactions with different HBGA types. Most human noroviruses bind HBGAs, while some strains were found to have minimal or no HBGA interactions. Here, we explain some possible structural constraints for several noroviruses that were found to bind poorly to HBGAs by using X-ray crystallography. We showed that one aspartic acid was flexible or positioned away from the fucose moiety of the HBGAs and this likely hindered binding, although other fucose-interacting residues were perfectly oriented. Interestingly, a neighboring loop also appeared to influence the loop hosting the aspartic acid. These new findings might explain why some human noroviruses bound HBGAs poorly, although further studies are required.

Download Full-text

The Distribution of Type 1 Chain ABH and Related Histo-Blood Group Antigens in Normal Cycling Human Endometrium

International Journal of Gynecological Pathology ◽

10.1097/00004347-199301000-00010 ◽

1993 ◽

Vol 12 (1) ◽

pp. 70-79 ◽

Cited By ~ 18

Author(s):

Vibeke Ravn ◽

Christence Stubbe Teglbjaerg ◽

Ulla Mandel ◽

Erik Dabelsteen

Keyword(s):

Blood Group ◽

Human Endometrium ◽

Blood Group Antigens

Download Full-text

Molecular basis of P[6] and P[8] major human rotavirus VP8* domain interactions with histo-blood group antigens

10.1101/512301 ◽

2019 ◽

Cited By ~ 2

Author(s):

Shenyuan Xu ◽

Yang Liu ◽

Ming Tan ◽

Weiming Zhong ◽

Dandan Zhao ◽

...

Keyword(s):

Nuclear Magnetic Resonance ◽

Magnetic Resonance ◽

Blood Group ◽

Molecular Basis ◽

Disease Burden ◽

Binding Domain ◽

Specific Cell ◽

Blood Group Antigens ◽

Nuclear Magnetic

AbstractInitial cell attachment of rotavirus (RV) to specific cell surface glycans, which is the essential first step in RV infection, is mediated by the VP8* domain of the spike protein VP4. Recently, human histo-blood group antigens (HBGAs) have been identified as ligands or receptors for human RV strains. RV strains in the P[4] and P[8] genotypes of the P[II] genogroup share common recognition of the Lewis b and H type 1 antigens, while P[6], which is one of the other genotypes in P[II], only recognizes the H type 1 antigen. The molecular basis of receptor recognition by the major human P[8] RVs remains unknown due to lack of experimental structural information. Here, we used nuclear magnetic resonance (NMR) titration experiments and NMR-derived high ambiguity driven docking (HADDOCK) methods to elucidate the molecular basis for P[8] VP8* recognition of the Leb and type 1 HBGAs and for P[6] recognition of H type 1 HBGAs. Unlike P[6] VP8* that recognizes H type 1 HGBAs in a binding surface composed of an α-helix and a β-sheet, referred as “βα binding domain”, the P[8] VP8* binds the type 1 HBGAs requiring the presence of the Lewis epitope in a previously undescribed pocket formed by two β-sheets, referred as “ββ binding domain”. The observation that P[6] and P[8] VP8* domains recognize different glycan structures at distinct binding sites supports the hypothesis that RV evolution is driven, at least in part, by selective pressure driven adaptation to HBGA structural diversity of their natural hosts living in the world. Recognition of the role that HBGAs play in driving RV evolution is essential to understanding RV diversity, host ranges, disease burden and zoonosis and to developing strategies to improve vaccines against RV infections.Author summaryRotaviruses (RV)s are the main cause of severe diarrhea in humans and animals. Significant advances in understanding RV diversity, evolution and epidemiology have been made after discovering that RVs recognize histo-blood group antigens (HBGAs) as host cell receptors. While different RV strains are known to have distinct binding preferences for HBGA receptors, the molecular basis in controlling strain-specific host ranges remains unclear. In this study, we used solution nuclear magnetic resonance to determine the molecular level details for interactions of the human P[8] and P[6] RV VP8* domains with their HBGA receptors. The distinct binding patterns observed between these major human RVs and their respective receptor ligands provides insight into the evolutionary relationships between different P[II] genotypes that ultimately determine host ranges, disease burden, zoonosis and epidemiology, which may impact future strategies for vaccine development against RVs.

Download Full-text

GII.13/21 Noroviruses Recognize Glycans with a Terminal β-Galactose via an Unconventional Glycan Binding Site

Journal of Virology ◽

10.1128/jvi.00723-19 ◽

2019 ◽

Vol 93 (15) ◽

Cited By ~ 2

Author(s):

Xin Cong ◽

Xiao-man Sun ◽

Jian-xun Qi ◽

Han-bo Li ◽

Wen-gang Chai ◽

...

Keyword(s):

Binding Site ◽

Blood Group ◽

Binding Mode ◽

Host Susceptibility ◽

Genetic Lineage ◽

Blood Group Antigens ◽

Human Noroviruses ◽

Control And Prevention ◽

P Domain ◽

Glycan Receptors

ABSTRACT Human noroviruses (huNoVs) recognize histo-blood group antigens (HBGAs) as host susceptibility factors. GII.13 and GII.21 huNoVs form a unique genetic lineage that emerged from mainstream GII NoVs via development of a new, nonconventional glycan binding site (GBS) that binds Lea antigen. This previous finding raised the question of whether the new GII.13/21 GBS really has such a narrow glycan binding spectrum. In this study, we provide solid phenotypic and structural evidence indicating that this new GBS recognizes a group of glycans with a common terminal β-galactose (β-Gal). First, we found that P domain proteins of GII.13/21 huNoVs circulating at different times bound three glycans sharing a common terminal β-Gal, including Lec, lactose, and mucin core 2. Second, we solved the crystal structures of the GII.13 P dimers in complex with Lec and mucin core 2, which showed that β-Gal is the major binding saccharide. Third, nonfat milk and lactose blocked the GII.13/21 P domain-glycan binding, which may explain the low prevalence of GII.13/21 viruses. Our data provide new insight into the host interactions and epidemiology of huNoVs, which would help in the control and prevention of NoV-associated diseases. IMPORTANCE Evidence from both phenotypic binding assay and structural study support the observed interactions of human noroviruses (huNoVs) with histo-blood group antigens (HBGAs) as receptors or attachment factors, affecting their host susceptibility. GII.13 and GII.21 genotypes form a unique genetic lineage that differs from the mainstream GII huNoVs in their unconventional glycan binding site. Unlike the previous findings that GII.13/21 genotypes recognize only Lea antigen, we found in this study that they can interact with a group of glycans with a common terminal β-Gal, including Lec, lactose, and mucin core 2. However, this wide glycan binding spectrum in a unique binding mode of the GII.13/21 huNoVs appears not to increase their prevalence, probably due to the existence of decoy glycan receptors in human gastrointestinal tract limiting their infection. Our findings shed light on the host interaction and epidemiology of huNoVs, which would impact the strategy of huNoV control and prevention.

Download Full-text