Immunosilencing a Highly Immunogenic Protein Trimerization Domain

Sliepen, Kwinten; Montfort, Thijs van; Melchers, Mark; Isik, Gözde; Sanders, Rogier W.

doi:10.1074/jbc.m114.620534

Cited by 69 publications

(89 citation statements)

References 51 publications

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“…The epitope of bNAb PGT151 is comprised of the gp120-gp41 interface of one protomer and glycans N611 and N637 of the adjacent protomer that were predicted to be complex-type glycans1260, consistent with results here. The other glycosites located in the gp41 region, N618 and N625, were found to be partially non-glycosylated, in keeping with earlier biochemical studies suggesting incomplete of occupancy glycosites in gp41 (refs 61, 62). …”

Section: Resultssupporting

confidence: 88%

Global site-specific N-glycosylation analysis of HIV envelope glycoprotein

et al. 2017

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HIV-1 envelope glycoprotein (Env) is the sole target for broadly neutralizing antibodies (bnAbs) and the focus for design of an antibody-based HIV vaccine. The Env trimer is covered by ∼90N-linked glycans, which shield the underlying protein from immune surveillance. bNAbs to HIV develop during infection, with many showing dependence on glycans for binding to Env. The ability to routinely assess the glycan type at each glycosylation site may facilitate design of improved vaccine candidates. Here we present a general mass spectrometry-based proteomics strategy that uses specific endoglycosidases to introduce mass signatures that distinguish peptide glycosites that are unoccupied or occupied by high-mannose/hybrid or complex-type glycans. The method yields >95% sequence coverage for Env, provides semi-quantitative analysis of the glycosylation status at each glycosite. We find that most glycosites in recombinant Env trimers are fully occupied by glycans, varying in the proportion of high-mannose/hybrid and complex-type glycans.

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Section: Resultssupporting

confidence: 88%

Global site-specific N-glycosylation analysis of HIV envelope glycoprotein

et al. 2017

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“…The Cal09 ST HA, PR8 ST HA, FM1 ST HA and cH6/1 ST HA feature a GCN4pII trimerization domain and a C-terminal Strep-Tag II. These substrates were chosen to avoid measuring reactivity against the immunogenic T4 foldon or the hexahistidine tag which are present on the PR8 HL HA antigen [29]. For ELISAs with PR8 ST HA, FM1 ST HA and cH6/1 ST HA an endpoint titer analysis was performed.…”

Section: Methodsmentioning

confidence: 99%

Vaccination with soluble headless hemagglutinin protects mice from challenge with divergent influenza viruses

Wohlbold

Nachbagauer

Margine

et al. 2015

Vaccine

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Current influenza virus vaccines provide solid protection from infection with viruses that are well matched with the vaccine strains. However, they do not protect efficiently against drifted or shifted strains. We developed an antigen based on the conserved stalk domain of the influenza virus hemagglutinin and tested its efficacy as a vaccine in a mouse virus challenge model. Although the antigen lacked the correct conformation of the native stalk domain and was not recognized by a panel of neutralizing stalk-reactive antibodies, it did induce considerable protection against H1N1, H5N1 and H6N1 challenge strains. Protection was enhanced when mice had pre-existing immunity against the stalk domain. Since pre-existing immunity is also present in the human population, we hypothesize that a similar antigen could show efficacy in humans as well.

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“…Motifs associated with cellular trafficking (localization, transport, secretion, and sequestration) are readily edited to modify where expression products go, and change interaction profiles with other proteins [14]. In addition to motifs that stabilize the structure of immunogens, such as trimerization ('foldon') [15][16][17][18] and dimerization [19,20] domains, motifs that interact with cellular processes for innate antiviral pathways could be used to enhance immunogenicity. While SLiMs in eukaryotic proteins have been discussed extensively, SLiM involvement in viral immunomodulation remains less thoroughly explored, and suggests new opportunities for use in engineered biotechnology applications.…”

Section: How Viruses Do More With Lessmentioning

confidence: 99%

Resources to Discover and Use Short Linear Motifs in Viral Proteins

Hraber

O’Maille

Silberfarb

et al. 2020

Trends in Biotechnology

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Viral proteins evade host immune function by molecular mimicry, often achieved by short linear motifs (SLiMs) of three to ten consecutive amino acids (AAs). Motif mimicry tolerates mutations, evolves quickly to modify interactions with the host, and enables modular interactions with protein complexes. Host cells cannot easily coordinate changes to conserved motif recognition and binding interfaces under selective pressure to maintain critical signaling pathways. SLiMs offer potential for use in synthetic biology, such as better immunogens and therapies, but may also present biosecurity challenges. We survey viral uses of SLiMs to mimic host proteins, and information resources available for motif discovery. As the number of examples continues to grow, knowledge management tools are essential to help organize and compare new findings.

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Immunosilencing a Highly Immunogenic Protein Trimerization Domain

Cited by 69 publications

References 51 publications

Global site-specific N-glycosylation analysis of HIV envelope glycoprotein

Global site-specific N-glycosylation analysis of HIV envelope glycoprotein

Vaccination with soluble headless hemagglutinin protects mice from challenge with divergent influenza viruses

Resources to Discover and Use Short Linear Motifs in Viral Proteins

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