Isolation and properties of the alpha-latrotoxin receptor.
The receptor protein of alpha-latrotoxin (alpha LTx, a neurotoxin with 'pure' presynaptic action isolated from black widow spider venom), was solubilized by Triton X-100 from bovine brain membranes and purified by affinity chromatography on alpha LTx-Sepharose. The purified receptor preparation contained four major polypeptides of molecular masses 200 (alpha), 160 (alpha'), 79 (beta) and 43 (gamma) kd according to SDS electrophoresis with molecular ratio alpha 1 alpha' 1 beta 2 gamma 2. The alpha- and alpha'-subunits are glycoproteins binding to wheat germ lectin and can be separated under non-denaturing conditions by anion exchange chromatography. Purified to homogeneity, both of them, though differing in the carbohydrate composition, retain the alpha LTx-binding activity and give closely related peptide maps. Anti-alpha antibodies recognize the alpha'-subunit as well. These results suggest that alpha LTx receptor is present in purified preparations in two very close forms containing the alpha- or alpha'-subunit. Beta and gamma proteins do not specifically bind alpha LTx and their physiological role is unclear. They form a complex with solubilized alpha- and alpha'-subunits independently of alpha LTx presence. The receptor proteins were purified to homogeneity by high performance gel filtration in the presence of SDS, their amino acid composition was determined.
Petrenko
A G
AG
Shemyakin Institute of Bioorganic Chemistry, USSR Academy of Sciences, Moscow.
Kovalenko
V A
VA
Shamotienko
O G
OG
Surkova
I N
IN
Tarasyuk
T A
TA
Ushkaryov YuA
Grishin
E V
EV
D000595:Amino Acid Sequence; D000818:Animals; D001923:Brain Chemistry; D002417:Cattle; D002540:Cerebral Cortex / Q000032:analysis*; D002846:Chromatography, Affinity; D002852:Chromatography, Ion Exchange; D007700:Kinetics; D008969:Molecular Sequence Data; D008970:Molecular Weight; D011487:Protein Conformation; D011950:Receptors, Cholinergic / Q000302:isolation & purification*; D018000:Receptors, Peptide
D011950:Receptors, Cholinergic; D018000:Receptors, Peptide; C034800:alpha-latrotoxin receptor