TY - JOUR
T1 - Polypeptide composition of the α-latrotoxin receptor
T2 - High affinity binding protein consists of a family of related high molecular weight polypeptides complexed to a low molecular weight protein
AU - Petrenko, A. G.
AU - Lazaryeva, V. D.
AU - Geppert, M.
AU - Tarasyuk, T. A.
AU - Moomaw, C.
AU - Khokhlatchev, A. V.
AU - Ushkaryov, Y. A.
AU - Slaughter, Clive A.
AU - Nasimov, I. V.
AU - Sudhof, T. C.
N1 - Copyright:
Copyright 2004 Elsevier B.V., All rights reserved.
PY - 1993/1/25
Y1 - 1993/1/25
N2 - α-Latrotoxin is a vertebrate neurotoxin from black widow spider venom that causes massive neurotransmitter release. In order to gain insight into the mechanism of action of α-latrotoxin, we have studied α-latrotoxin-binding proteins from bovine and rat brain. Proteins purified by affinity chromatography on immobilized α-latrotoxin were investigated. Two sets of proteins were isolated: 1) three polypeptides of Mr 79,000, 65,000, and 43,000 that were eluted from immobilized a-latrotoxin by increasing KCl concentrations in the presence of Ca2+, and 2) a family of related proteins ranging in molecular weight from 160,000 to 220,000 and a low molecular weight component of Mr 29,000 that were eluted from immobilized α-latrotoxin only after removal of Ca2+. Amino acid sequences of these proteins demonstrated that all of these proteins represent novel proteins except for the Mr 65,000 polypeptide, which is identical with synaptotagmin (Petrenko, A. G., Perin, M. S., Davletov, B. A., Ushkaryov, Y. A., Geppert, M., and Südhof, T. C. (1991) Nature 353, 65-68). Surprisingly, the Mr 79,000 and 43,000 proteins were also found in tissues insensitive to α-latrotoxin action. Since these proteins do not bind 125I-α-latrotoxin with high affinity, their purification probably is not physiologically significant. On the other hand, the fractions containing the Mr 160,000-220,000 and 29,000 polypeptides bound α-latrotoxin with high affinity. Sucrose gradient centrifugations and anion exchange chromatography suggested that most of the Mr 160,000-220,000 proteins were complexed with the Mr 29,000 protein. α-Latrotoxin binding correlated with the presence of the Mr 160,000-220,000 proteins and Mr 29,000 polypeptide, and α-latrotoxin formed stable complexes with the Mr 160,000-220,000 proteins. Accordingly, the α-latrotoxin receptor consists of a high molecular weight protein (Mr 160,000-220,000) that is complexed with one or several copies of an Mr 29,000 polypeptide. In addition, the receptor is found in a less tight association with synaptotagmin but not with other polypeptides.
AB - α-Latrotoxin is a vertebrate neurotoxin from black widow spider venom that causes massive neurotransmitter release. In order to gain insight into the mechanism of action of α-latrotoxin, we have studied α-latrotoxin-binding proteins from bovine and rat brain. Proteins purified by affinity chromatography on immobilized α-latrotoxin were investigated. Two sets of proteins were isolated: 1) three polypeptides of Mr 79,000, 65,000, and 43,000 that were eluted from immobilized a-latrotoxin by increasing KCl concentrations in the presence of Ca2+, and 2) a family of related proteins ranging in molecular weight from 160,000 to 220,000 and a low molecular weight component of Mr 29,000 that were eluted from immobilized α-latrotoxin only after removal of Ca2+. Amino acid sequences of these proteins demonstrated that all of these proteins represent novel proteins except for the Mr 65,000 polypeptide, which is identical with synaptotagmin (Petrenko, A. G., Perin, M. S., Davletov, B. A., Ushkaryov, Y. A., Geppert, M., and Südhof, T. C. (1991) Nature 353, 65-68). Surprisingly, the Mr 79,000 and 43,000 proteins were also found in tissues insensitive to α-latrotoxin action. Since these proteins do not bind 125I-α-latrotoxin with high affinity, their purification probably is not physiologically significant. On the other hand, the fractions containing the Mr 160,000-220,000 and 29,000 polypeptides bound α-latrotoxin with high affinity. Sucrose gradient centrifugations and anion exchange chromatography suggested that most of the Mr 160,000-220,000 proteins were complexed with the Mr 29,000 protein. α-Latrotoxin binding correlated with the presence of the Mr 160,000-220,000 proteins and Mr 29,000 polypeptide, and α-latrotoxin formed stable complexes with the Mr 160,000-220,000 proteins. Accordingly, the α-latrotoxin receptor consists of a high molecular weight protein (Mr 160,000-220,000) that is complexed with one or several copies of an Mr 29,000 polypeptide. In addition, the receptor is found in a less tight association with synaptotagmin but not with other polypeptides.
UR - http://www.scopus.com/inward/record.url?scp=0027492728&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=0027492728&partnerID=8YFLogxK
M3 - Article
C2 - 8420960
AN - SCOPUS:0027492728
SN - 0021-9258
VL - 268
SP - 1860
EP - 1867
JO - Journal of Biological Chemistry
JF - Journal of Biological Chemistry
IS - 3
ER -