Characterization of a rabbit serum raised against a botulinum toxin type A binding protein from presynaptic plasma membranes from Torpedo electric organ.
Botulinum neurotoxin type A blocks acetylcholine release from the peripheral nervous system. We have previously described a putative botulinum neurotoxin type A receptor of presynaptic plasma membranes from Torpedo. The electric organ of Torpedo, which is largely enriched in cholinergic nerve endings, is homologous to the neuromuscular junction, allowing us to isolate large scale of presynaptic components. In order to characterize this protein we have raised a polyclonal antibody (a-P140) against this receptor. The antiserum a-P140 recognizes a 140,000 mol. wt band in non-reducing conditions and an 80,000 band in reducing conditions. The immunohistochemistry assay reveals the P140 protein on the ventral face of the electrocytes where the nerve terminals are localized. Moreover, a-P140 antiserum recognizes the P140-BoNT/A complex after binding and cross-linking experiments. In addition, we have immunoprecipitated an in vitro translated product which is closely coincident in mol. wt to the 80,000 band of the receptor.
Canals
J M
JM
Departament de Biología Cellular i Anatomía Patològica, Hospital de Bellvitge, Facultat de Medicina, Universitat de Barcelona, Spain.
Arribas
M
M
Ruiz-Avila
L
L
Blasi
J M
JM
Marsal
J
J
Solsona
C
C
D000818:Animals; D001427:Bacterial Toxins / Q000378:metabolism*; D015153:Blotting, Western; D002135:Calcium-Binding Proteins; D002352:Carrier Proteins / Q000276:immunology* / Q000378:metabolism; D003016:Clostridium perfringens; D003432:Cross-Linking Reagents; D004557:Electric Organ / Q000378:metabolism*; D007150:Immunohistochemistry; D066298:In Vitro Techniques; D008970:Molecular Weight; D009411:Nerve Endings / Q000378:metabolism; D010275:Parasympathetic Nervous System / Q000378:metabolism; D011233:Precipitin Tests; D011485:Protein Binding; D014176:Protein Biosynthesis; D012333:RNA, Messenger / Q000096:biosynthesis; D011817:Rabbits / Q000276:immunology; D017661:Receptors, Presynaptic / Q000378:metabolism; D012156:Reticulocytes / Q000378:metabolism; D013570:Synaptic Membranes / Q000378:metabolism*; D014101:Torpedo / Q000378:metabolism*; D010738:Type C Phospholipases
D001427:Bacterial Toxins; D002135:Calcium-Binding Proteins; D002352:Carrier Proteins; D003432:Cross-Linking Reagents; D012333:RNA, Messenger; D017661:Receptors, Presynaptic; D010738:Type C Phospholipases; C040649:alpha toxin, Clostridium perfringens