Isolation and characterization of extremely minor gangliosides, GM1b and GD1 alpha, in adult bovine brains as developmentally regulated antigens.
In addition to ganglioside GM1b, an unusual and extremely minor ganglioside, GD1 alpha, was efficiently isolated from bovine brain by combination of Q-Sepharose and Iatrobeads column chromatographies. In the course of purification steps, the presence of the sialidase-labile ganglioside was proved by a highly sensitive TLC/enzyme-immunostaining method. The structure was characterized by gas-liquid chromatography, permethylation study, sialidase degradation, immunostaining with specific antibodies, fast atom bombardment-mass spectrometry, and proton magnetic resonance spectrometry. The content of the ganglioside was very small (0.016%) in the total gangliosides. This finding suggests that a synthetic pathway of asialo GM1----GM1b----GD1 alpha may exist in mammalian brains. A monoclonal antibody NA-6 that was obtained by immunizing mice with purified GM1b reacted specifically with GM1b but showed no cross-reactivity with other structurally related gangliosides such as GM1a, GD1a, and so on. Using the method of TLC/immunostaining with NA-6, GM1b was found to be strongly expressed during embryonic days 14-17 in chick brains. Thus, it is assumed that extremely minor gangliosides like GM1b and GD1 alpha found in adult brains are characterized as embryonic molecules.
Department of Biochemistry, University of Shizuoka, School of Pharmaceutical Sciences, Japan.
Y
Y
Hirabayashi
A
A
Hyogo
T
T
Nakao
K
K
Tsuchiya
Y
Y
Suzuki
M
M
Matsumoto
K
K
Kon
S
S
Ando
D000818:Animals; D001921:Brain / Q000196:embryology / Q000254:growth & development*; D001923:Brain Chemistry; D002236:Carbohydrate Conformation; D002240:Carbohydrate Sequence; D002642:Chick Embryo; D002845:Chromatography; D002855:Chromatography, Thin Layer; D005677:G(M1) Ganglioside / Q000031:analogs & derivatives* / Q000302:isolation & purification; D006801:Humans; D007124:Immunoenzyme Techniques; D007231:Infant, Newborn; D009682:Magnetic Resonance Spectroscopy; D013058:Mass Spectrometry; D008969:Molecular Sequence Data; D015394:Molecular Structure
C064020:ganglioside GD1alpha; D005677:G(M1) Ganglioside; C029624:ganglioside M1b