Fig. 1

Analysis of N-linked glycosylation in CedV F protein. a Schematic of localization of potential N-glycans. N-glycosylation mutants are named g1–g6. Black triangles indicate potential N-glycosylation sites within the F1 and F2 subunits. TM: transmembrane domain; CD: cytoplasmic domain; b PNGase F digest of CedV F protein. MDCK cells expressing CedV F protein were metabolically labeled for 15 min (pulse) and then incubated for 2 h in serum-free nonradioactive medium (chase). After immunoprecipitation of F proteins from cell lysates, samples were suspended in sample buffer and then treated with N-glycosidase F (PNGase F) according to the instructions of the manufacturer, or left untreated. After separation on a 12% SDS-gel under reducing conditions, samples were analyzed by autoradiography; c Proteolytic processing of CedV F and F mutants. MDCK cells expressing F proteins are metabolically labeled as described above. After immunoprecipitation of F proteins from cell lysates and separation on a 12% SDS-gel under reducing conditions, samples were analyzed by autoradiography. Molecular masses of marker proteins are indicated. n = 2. Stars indicate the F₂ subunit