Structure and receptor recognition by the Lassa virus spike complex

Katz, Michael J.; Weinstein, Jonathan; Eilon-Ashkenazy, Maayan; Cohen-Dvashi, Hadas; Elad, Nadav; Fleishman, Sarel J.; Diskin, Ron

doi:10.1038/s41586-022-04429-2

Cited by 44 publications

(86 citation statements)

References 66 publications

(86 reference statements)

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“…It is possible that the conformational properties of matriglycan are length-dependent and that a threshold length is required to adopt a recognition conformation. This possibility is supported by a recent cryo-EM structure of the LASV Spike protein that indicate it can accommodate a matriglycan of at least 13 monosaccharides in a defined spiral-like conformation 63 . It is also conceivable that longer matriglycans can provide a scaffold for multivalent interactions with LG domain-containing proteins resulting in high avidity binding 64 .…”

Section: Discussionmentioning

confidence: 86%

Cell surface glycan engineering reveals that matriglycan alone can recapitulate dystroglycan binding and function

et al. 2022

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Abstractα-Dystroglycan (α-DG) is uniquely modified on O-mannose sites by a repeating disaccharide (-Xylα1,3-GlcAβ1,3-)n termed matriglycan, which is a receptor for laminin-G domain-containing proteins and employed by old-world arenaviruses for infection. Using chemoenzymatically synthesized matriglycans printed as a microarray, we demonstrate length-dependent binding to Laminin, Lassa virus GP1, and the clinically-important antibody IIH6. Utilizing an enzymatic engineering approach, an N-linked glycoprotein was converted into a IIH6-positive Laminin-binding glycoprotein. Engineering of the surface of cells deficient for either α-DG or O-mannosylation with matriglycans of sufficient length recovers infection with a Lassa-pseudovirus. Finally, free matriglycan in a dose and length dependent manner inhibits viral infection of wildtype cells. These results indicate that matriglycan alone is necessary and sufficient for IIH6 staining, Laminin and LASV GP1 binding, and Lassa-pseudovirus infection and support a model in which it is a tunable receptor for which increasing chain length enhances ligand-binding capacity.

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Section: Discussionmentioning

confidence: 86%

Cell surface glycan engineering reveals that matriglycan alone can recapitulate dystroglycan binding and function

et al. 2022

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“…The absence or reduction of matriglycan can cause various forms of muscular dystrophy with or without abnormalities in brain development ( 7, 8 ). Additionally, matriglycan is a receptor for Lassa fever virus (LASV) and other Old-World mammarenaviruses ( 9, 10 ). Matriglycan polymer length appears to be cell-type specific and is physiologically regulated during development and regeneration ( 11 ) but can be pathologically decreased by the expression of LASV or the related lymphocytic choriomeningitis virus spike glycoproteins ( 12 ).…”

Section: Introductionmentioning

confidence: 99%

Structure and mechanism of LARGE1 matriglycan polymerase

Joseph

Schnicker

et al. 2022

Preprint

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Matriglycan is a linear polysaccharide of alternating xylose and glucuronate that binds extracellular matrix proteins and acts as a receptor for Lassa fever virus. LARGE1 synthesizes matriglycan on dystroglycan and mutations in LARGE1 cause muscular dystrophy with abnormal brain development. However, the mechanism of matriglycan polymerization by LARGE1 is unknown. Here, we report the cryo-EM structure of LARGE1. We show that LARGE1 functions as a dimer to polymerize matriglycan by alternating activities between the xylose transferase domain on one protomer and the glucuronate transferase domain on the other protomer. Biochemical analyses using a recombinant Golgi form of dystroglycan reveal that LARGE1 polymerizes matriglycan processively. Our results provide mechanistic insights into LARGE1 function and may facilitate novel therapeutic strategies for treating neuromuscular disorders or arenaviral infections.One-Sentence SummaryDimeric LARGE1 processively polymerizes matriglycan on dystroglycan using orthogonal active sites on alternate protomers.

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“…When compared to the mature closed structures (8, 12, 20), D5 would be obstructed from binding at the apex ( Fig. 6C ).…”

Section: Resultsmentioning

confidence: 99%

“…The surface of LASV virions is covered by the trimeric type 1-fusion glycoprotein complex (GPC) (6,7), the only antigen available for virus-neutralizing antibodies. Each protomer of the GPC consists of a receptor-binding GP1 subunit, a transmembrane-spanning GP2 subunit, and the stably-associated signal peptide (SSP) (8)(9)(10)(11)(12). The highly glycosylated GPC induces weak and inconsistent immune responses in both natural infection and vaccination (13)(14)(15)(16)(17).…”

Section: Introductionmentioning

confidence: 99%

Prefusion-Stabilized Lassa Virus Trimer Identifies Neutralizing Nanobodies and Reveals an Apex-Situated Site of Vulnerability

Gorman

Cheung

Duan

et al. 2022

Preprint

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Lassa virus (LASV) is responsible for 100,000-300,000 zoonotic infections annually and poses a threat to public health. Development of antibody-based therapeutics or vaccines has been challenging because neutralizing antibodies – even among Lassa hemorrhagic fever survivors – are generally of low titer, and the target of neutralizing antibodies, the trimeric glycoprotein complex (GPC), a type 1-fusion machine with GP1 and GP2 subunits, has been difficult to produce. Here, we use structure-based design to obtain a soluble LASV GPC by engineering an inter-protomer disulfide (R207GCGP1-L326CGP2) and appending the T4-fibritin trimerization domain. We verified the antigenicity of this prefusion-stabilized LASV GPC against a panel of human antibodies and used electron microscopy (EM) to confirm its trimeric association. We panned the prefusion-stabilized LASV GPC against single domain ‘nanobody’ libraries and identified one of camel origin, which we named D5, which bound GPC with 27 nM affinity and neutralized the Josiah strain of LASV with an IC50 of 12 µg/ml when formatted into a bivalent IgG2a context. The cryo-EM structure of a ternary complex of the D5 nanobody, the antigen-binding fragment of human antibody 8.11G, and LASV GPC revealed D5 to recognize a site-of-vulnerability at the trimer apex. The recognized site appeared to be specific to GPC lacking cleavage of between GP1 and GP2 subunits. Collectively, our findings suggest that GPC-cleavage intermediates may be targets for LASV neutralization and define an apex-situated site of vulnerability for vaccine development.SignificanceLassa virus (LASV) infection is expanding outside its traditionally endemic areas in West Africa, posing a biothreat to the world. LASV-neutralizing antibodies, moreover, have proven difficult to elicit. To gain insight into requirements for antibody-mediated neutralization of LASV, we developed a prefusion-stabilized LASV glycoprotein trimer (GPC), panned it against phage libraries comprised single-domain antibodies or nanobodies from shark and camel, and identified one, D5, which – when placed into bivalent IgG2a context – could neutralize LASV. Cryo-EM analysis revealed D5 to recognize a cleavage-dependent site-of-vulnerability at the trimer apex. We propose this apex-situated site to be an attractive target for LASV vaccine and therapeutic development.

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Structure and receptor recognition by the Lassa virus spike complex

Abstract: Articlestructure of the LASV spike complex, which provides fundamental information regarding the SSP, receptor recognition and the stability of this trimeric complex.

Cited by 44 publications

References 66 publications

Cell surface glycan engineering reveals that matriglycan alone can recapitulate dystroglycan binding and function

Cell surface glycan engineering reveals that matriglycan alone can recapitulate dystroglycan binding and function

Structure and mechanism of LARGE1 matriglycan polymerase

Prefusion-Stabilized Lassa Virus Trimer Identifies Neutralizing Nanobodies and Reveals an Apex-Situated Site of Vulnerability

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