Ganglioside binding proteins of calf brain with ubiquitin-like N-terminals.
Two ganglioside-associated protein components I and II have been isolated from crude ganglioside preparations of calf brain by DEAE-Sephadex ion-exchange chromatography. Both components exhibited binding capacity in aqueous media for gangliosides of the 'ganglio' series but not for neutral glycosphingolipids (polyglycosylceramides) and only a low capacity for sialosylparagloboside. Each protein bound individual gangliosides with different efficiency. Upon prolonged incubation of component I with gangliosides, complexes with high (30:1) and low (6:1) glycolipid/protein molar ratios were formed. The latter but not the former complex was able to penetrate Sephadex G-200 beads. Both components inhibited plating efficiency of cultured mouse N2a neuroblastoma cells. The molecular masses of components I and II were determined by SDS/PAGE to be 11-12 kDa and 28 kDa, respectively. Carbohydrates (fucose, mannose, galactose, N-acetylglucosamine, N-acetylgalactosamine, and some sialic acid) were found only in component II. When examined by reverse-phase HPLC each component separated into two major closely migrating peaks which were subsequently examined by Edman degradation. Amino acid sequences of the N-terminal portions of three of these peaks (one peak from component I and both peaks from component II) showed, as far as the sequences were established, identity with the sequence of ubiquitin. It is hypothesized that the proteins may be instrumental in intracellular trafficking of gangliosides.
Zdebska
E
E
Zaklad Biochemii, Instytut Hematologii, Warszawa, Poland.
Antoniewicz
J
J
Nilsson
B
B
Sandhoff
K
K
Fürst
W
W
Janik
P
P
Kościelak
J
J
D000595:Amino Acid Sequence; D000818:Animals; D001923:Brain Chemistry; D002240:Carbohydrate Sequence; D002241:Carbohydrates / Q000032:analysis; D002352:Carrier Proteins / Q000737:chemistry / Q000302:isolation & purification* / Q000378:metabolism; D002417:Cattle; D002455:Cell Division / Q000187:drug effects; D002851:Chromatography, High Pressure Liquid; D005732:Gangliosides / Q000378:metabolism*; D051379:Mice; D008969:Molecular Sequence Data; D008970:Molecular Weight; D009419:Nerve Tissue Proteins / Q000737:chemistry / Q000302:isolation & purification* / Q000378:metabolism; D010446:Peptide Fragments / Q000737:chemistry; D017386:Sequence Homology, Amino Acid; D013109:Sphingomyelins / Q000378:metabolism; D014407:Tumor Cells, Cultured; D014452:Ubiquitins / Q000737:chemistry*
D002241:Carbohydrates; D002352:Carrier Proteins; D005732:Gangliosides; D009419:Nerve Tissue Proteins; D010446:Peptide Fragments; D013109:Sphingomyelins; D014452:Ubiquitins