The nuclear pore complex protein p62 is one of several sialic acid-containing proteins of the nuclear envelope.
While investigating the glycosylation of nuclear envelope proteins of neuroblastoma cells, we found several proteins that bound the sialic acid-specific Sambucus nigra agglutinin. The strongest signals were obtained for proteins with apparent molecular masses of 66 and 180 kDa. The specificity of the lectin binding was checked by acylneuraminyl hydrolase treatment of nuclear envelope proteins, which prohibited S. nigra agglutinin binding. Digestion of nuclear envelope proteins with the N-glycosidase F revealed that sialic acid was N-glycosidically linked to the 180-kDa protein and very probably O-glycosidically linked to the 66-kDa protein. Upon extraction, the latter behaved like the nucleoporin p62 in that it was partly extracted by high ionic strength buffers, could not be solubilized by nonionic detergent, and was completely removed from the nuclear envelope with urea. Two-dimensional gel electrophoretic comparison showed that the S. nigra agglutinin-binding protein and p62 have an identical isoelectric point of about 5.0 and an identical apparent molecular mass of 66 kDa. This, together with the binding of the anti-nucleoporin antibody, demonstrated the identity of the 66-kDa sialoprotein and p62. S. nigra agglutinin inhibits nuclear protein transport in neuroblastoma cells, strongly suggesting a functional significance of sialylation of p62.
Emig
S
S
Arbeitsgruppe Neurochemie, Institut für Biochemie, Freie Universität Berlin, Federal Republic of Germany.
Schmalz
D
D
Shakibaei
M
M
Buchner
K
K
D000373:Agglutinins / Q000494:pharmacology; D000818:Animals; D001692:Biological Transport; D015180:Electrophoresis, Gel, Two-Dimensional; D051379:Mice; D008970:Molecular Weight; D019158:N-Acetylneuraminic Acid; D009685:Nuclear Envelope / Q000737:chemistry*; D009687:Nuclear Proteins / Q000032:analysis* / Q000378:metabolism; D012794:Sialic Acids / Q000032:analysis*; D014407:Tumor Cells, Cultured
D000373:Agglutinins; D009687:Nuclear Proteins; D012794:Sialic Acids; D019158:N-Acetylneuraminic Acid