The Diversity of Yellow-Related Proteins in Sand Flies (Diptera: Psychodidae)

Šíma, Michal; Novotný, Marián; Pravda, Lukáš; Sumova, Petra; Rohoušová, Iva; Volf, Petr

doi:10.1371/journal.pone.0166191

Cited by 12 publications

(11 citation statements)

References 59 publications

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“…longipalpis salivary proteins and their biological roles have been well studied 4,20 ; however, the sugars that modify these proteins have not been characterised in detail. Most work on sand fly salivary glycans comes from in silico analyses 13-15,17,18,25 and lectin blotting. They were first reported by Volf et al 19 , who used lectins to detect mannosylated N -type glycans.…”

Section: Discussionmentioning

confidence: 99%

Insights into the salivaryN-glycome ofLutzomyia longipalpis, vector of visceral leishmaniasis

Mondragón-Shem

Wongtrakul-Kish

Kozak

et al. 2020

Preprint

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24During Leishmania transmission sand flies inoculate parasites and saliva into the skin of 25 vertebrates. Saliva has anti-haemostatic and anti-inflammatory activities that evolved to facilitate 26 bloodfeeding, but also modulate the host's immune responses. Sand fly salivary proteins have 27 been extensively studied, but the nature and biological roles of protein-linked glycans remain 28 overlooked. Here, we characterised the profile of N-glycans from the salivary glycoproteins of 29 Lutzomyia longipalpis, vector of visceral leishmaniasis in the Americas. In silico predictions 30 suggest half of Lu. longipalpis salivary proteins may be N-glycosylated. SDS-PAGE coupled to LC-31 MS analysis of sand fly saliva, before and after enzymatic deglycosylation, revealed several 32 candidate glycoproteins. To determine the diversity of N-glycan structures in sand fly saliva, 33 enzymatically released sugars were fluorescently tagged and analysed by HPLC, combined with 34 highly sensitive LC-MS/MS, MALDI-TOF-MS, and exoglycosidase treatments. We found that the 35 N-glycan composition of Lu. longipalpis saliva mostly consists of oligomannose sugars, with 36Man5GlcNAc2 being the most abundant, and a few hybrid-type species. Interestingly, some 37 glycans appear modified with a group of 144 Da, whose identity has yet to be confirmed. Our 38 work presents the first detailed structural analysis of sand fly salivary glycans. 39 40 41 42 43 44 45 72There are some reports describing the presence of salivary glycoproteins in sand flies through in 73 silico and blotting analyses [13][14][15][16][17][18][19] ; however, to our knowledge no detailed structural studies have 74 been published to date. Therefore, we set out to identify the salivary glycoproteins in the sand fly 75 vector species Lutzomyia longipalpis, and structurally characterise their N-glycan conjugates. We 76 further discuss their implications for insect bloodfeeding as well as vector-host interactions. 77 78 Results 79 Identification of Lutzomyia longipalpis salivary glycoproteins.80 To determine the degree of N-glycosylation, an in silico analysis was carried out on 42 salivary 81 proteins previously reported in Lu. longipalpis 4,20 to predict protein N-glycosylation sites using the 82 NetNGlyc server (http://www.cbs.dtu.dk/services/NetNGlyc/). This revealed 48% of the 83 commonly known salivary proteins contain conventional N-glycosylation sites (Supplementary 84 those without recognizable glycosylation sequons (as determined by NetNGlyc), obtaining a final 109 list of 43 potentially N-glycosylated protein candidates (Supplementary Table S2). Fourteen of 110 these potential glycoproteins were also identified in our initial in silico analysis (Supplementary 111 Table S1), including LJM11, LJM111 and LJL143, which have been proposed as potential vaccine 112 components against Leishmania infection 4 . Using the InterProScan tool to identify conserved 113 protein domains, family distributions ( Supplementary Fig. S4) show five of the candidates 114 belonging ...

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Section: Discussionmentioning

confidence: 99%

Insights into the salivaryN-glycome ofLutzomyia longipalpis, vector of visceral leishmaniasis

Mondragón-Shem

Wongtrakul-Kish

Kozak

et al. 2020

Preprint

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“…longipalpis saliva specific IgG antibodies, in individuals living in CL or VL endemic areas, respectively [32,49]. Differences in salivary proteins from various sand fly species could explain these discrepancies [58–60,65]. …”

Section: Discussionmentioning

confidence: 99%

Human cellular and humoral immune responses to Phlebotomus papatasi salivary gland antigens in endemic areas differing in prevalence of Leishmania major infection

et al. 2017

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BackgroundSand fly saliva compounds are able to elicit specific immune responses that have a significant role in Leishmania parasite establishment and disease outcome. Characterizing anti-saliva immune responses in individuals living in well defined leishmaniasis endemic areas would provide valuable insights regarding their effect on parasite transmission and establishment in humans.Methodology/Principal findingsWe explored the cellular and humoral immune responses to Phlebotomus (P.) papatasi salivary gland extracts (SGE) in individuals living in cutaneous leishmaniasis (CL) old or emerging foci (OF, EF). OF was characterized by a higher infection prevalence as assessed by higher proportions of leishmanin skin test (LST) positive individuals compared to EF. Subjects were further subdivided into healed, asymptomatic or naïve groups. We showed anti-SGE proliferation in less than 30% of the individuals, regardless of the immune status, in both foci. IFN-γ production was higher in OF and only observed in immune individuals from OF and naïve subjects from EF. Although IL-10 was not detected, addition of anti-human IL-10 antibodies revealed an increase in proliferation and IFN-γ production only in individuals from OF. The percentage of seropositive individuals was similar in immune and naïves groups but was significantly higher in OF. No correlation was observed between anti-saliva immune responses and LST response. High anti-SGE-IgG responses were associated with an increased risk of developing ZCL. No differences were observed for anti-SGE humoral or cellular responses among naïve individuals who converted or not their LST response or developed or not ZCL after the transmission season.Conclusions/SignificanceThese data suggest that individuals living in an old focus characterized by a frequent exposure to sand fly bites and a high prevalence of infection, develop higher anti-saliva IgG responses and IFN-γ levels and a skew towards a Th2-type cellular response, probably in favor of parasite establishment, compared to those living in an emerging focus.

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“…longipalpis salivary proteins and their biological roles have been well studied 4 , 20 ; however, the sugars that modify these proteins have not been characterised in detail. Most work on sand fly salivary glycans comes from in silico analyses 13 – 15 , 17 , 18 , 25 and lectin blotting. They were first reported by Volf et al 19 , who used lectins to detect mannosylated N -type glycans.…”

Section: Discussionmentioning

confidence: 99%

Insights into the salivary N-glycome of Lutzomyia longipalpis, vector of visceral leishmaniasis

Mondragón-Shem

Wongtrakul-Kish

Kozak

et al. 2020

Sci Rep

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During Leishmania transmission sand flies inoculate parasites and saliva into the skin of vertebrates. Saliva has anti-haemostatic and anti-inflammatory activities that evolved to facilitate bloodfeeding, but also modulate the host's immune responses. Sand fly salivary proteins have been extensively studied, but the nature and biological roles of protein-linked glycans remain overlooked. Here, we characterised the profile of N-glycans from the salivary glycoproteins of Lutzomyia longipalpis, vector of visceral leishmaniasis in the Americas. in silico predictions suggest half of Lu. longipalpis salivary proteins may be N-glycosylated. SDS-PAGE coupled to LC-MS analysis of sand fly saliva, before and after enzymatic deglycosylation, revealed several candidate glycoproteins. to determine the diversity of N-glycan structures in sand fly saliva, enzymatically released sugars were fluorescently tagged and analysed by HPLC, combined with highly sensitive LC-MS/MS, MALDI-TOF-MS, and exoglycosidase treatments. We found that the N-glycan composition of Lu. longipalpis saliva mostly consists of oligomannose sugars, with Man 5 GlcnAc 2 being the most abundant, and a few hybrid-type species. Interestingly, some glycans appear modified with a group of 144 Da, whose identity has yet to be confirmed. Our work presents the first detailed structural analysis of sand fly salivary glycans. Sand flies are small insects that can transmit bacteria and viruses 1,2 , but are known mainly as vectors of leishmaniasis, a disease that threatens 350 million people worldwide 3. When female sand flies feed, they inject a saliva comprised of molecules that facilitate the ingestion of blood, and modulate the host immune system and pathogen transmission 4-6. These effects have led researchers to explore the potential of insect salivary molecules as markers of biting exposure 5,7 (to determine risk of disease), or even as components of vaccines against leishmaniasis 8. Of the many types of molecules that make up saliva, most research has focused on the proteins; here, we have investigated the glycans that modify these proteins. In most eukaryotic cells, the addition of glycans to proteins is a highly conserved and diverse post-translational modification. The most common types of protein-linked glycans are N-linked (attached to asparagine residues in the sequon Asn-X-Thr/Ser), and O-linked (attached to serine or threonine residues). Glycoconjugates display a wide range of biological roles, from organism development to immune system functions against pathogens 9. One study has addressed the types and roles of glycans in insects using the model fruit fly, Drosophila melanogaster. In this species, biological functions have been attributed to different glycan classes, such as morphology and locomotion (N-linked glycans), or cell interaction and signalling (O-linked glycans) 10 .

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The Diversity of Yellow-Related Proteins in Sand Flies (Diptera: Psychodidae)

Cited by 12 publications

References 59 publications

Insights into the salivaryN-glycome ofLutzomyia longipalpis, vector of visceral leishmaniasis

Insights into the salivaryN-glycome ofLutzomyia longipalpis, vector of visceral leishmaniasis

Human cellular and humoral immune responses to Phlebotomus papatasi salivary gland antigens in endemic areas differing in prevalence of Leishmania major infection

Insights into the salivary N-glycome of Lutzomyia longipalpis, vector of visceral leishmaniasis

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