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Figure 5 | Virology Journal

Figure 5

From: Characterization of the Lassa virus GP1 ectodomain shedding: implications for improved diagnostic platforms

Figure 5

Glycosylation profile of GPC and sGP1-RRAA-FLAG-derived secreted GP1 assessed by digestion with endo- and exo-glycosidases, and by lectin binding profiles. Immunoprecipitated secreted GP1 from LASV GPC expression (A) or sGP1-RRAA-FLAG expression (B) was subjected to digestion with PNGase F (lane 2), Endo H (lane 3), or neuraminidase (lane 4). Untreated glycoprotein controls were similarly processed, as outlined in methods, but enzymes were not added to reactions (lane 1). Proteins were subjected to SDS-PAGE and western blot analysis with biotinylated anti-GP1 mAb L52-74-7A and SA-HRP. Molecular weights of fully glycosylated GP1 (42 KDa) and the completely deglycosylated form of the protein (22 KDa), are indicated. (C) GNA lectin binding profile on immunoprecipitated GP1 from GPC expression (lane 1) or sGP1-RRAA-FLAG (lane 3). Control GNA binding for each reaction was performed on CpY (lanes 2 and 4). The heavy chain of mAb L52-74-7A reacted with GNA, and is indicated (mAb HC). Secreted GP1 from GPC expression reacted significantly stronger with GNA (lane 1) than with sGP1-RRAA-FLAG (lane 3). (D) Complete lectin panel binding profile on secreted GP1 from GPC or sGP1-RRAA-FLAG.

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