Identification and purification of a very high affinity binding protein for toxic phospholipases A2 in skeletal muscle.
Oxyuranus scutellatus toxin 1 (OS1) and toxin 2 (OS2) are two monochain phospholipases A2 isolated from the venom of Taipan. Their iodinated derivatives have been used to characterize phospholipase A2 receptors on rabbit skeletal muscle cells in culture. Both ligands recognize one family of binding sites on myotube membranes with a Bmax of 1.9 to 2.2 pmol/mg of protein and dissociation constant values of 7.4 pM for 125I-OS2 and 38 pM for 125I-OS1. Other snake venom phospholipases A2 are able to inhibit 125I-OS2 binding to the muscle receptor. Competition experiments with these unlabeled phospholipases A2 define a pharmacological profile of the muscle receptor very different from the previously described pharmacological profile of the neuronal phospholipase A2 receptors. The number of 125I-OS2 receptors in skeletal muscle cells increases during in vitro cell maturation but there is no clear relation between the increase of Bmax and the fusion of myoblasts into myotubes. The phospholipase A2 binding protein from myotubes has been identified both by cross-linking experiments and by purification studies. It is composed of only one subunit of Mr 180,000.
Lambeau
G
G
Institut de Pharmacologie Moléculaire et Cellulaire, UPR 411 du Centre National de la Recherche Scientifique, Valbonne, France.
Schmid-Alliana
A
A
Lazdunski
M
M
Barhanin
J
J
D000345:Affinity Labels; D000818:Animals; D001667:Binding, Competitive; D002454:Cell Differentiation; D002462:Cell Membrane / Q000032:analysis; D002478:Cells, Cultured; D002793:Cholic Acids; D002845:Chromatography; D003432:Cross-Linking Reagents; D004546:Elapid Venoms / Q000032:analysis; D004591:Electrophoresis, Polyacrylamide Gel; D008970:Molecular Weight; D024510:Muscle Development; D009132:Muscles / Q000032:analysis*; D010740:Phospholipases / Q000378:metabolism*; D010741:Phospholipases A / Q000378:metabolism*; D054467:Phospholipases A2; D011817:Rabbits; D011956:Receptors, Cell Surface / Q000032:analysis* / Q000378:metabolism
D000345:Affinity Labels; D002793:Cholic Acids; D003432:Cross-Linking Reagents; D004546:Elapid Venoms; D011956:Receptors, Cell Surface; D010740:Phospholipases; D010741:Phospholipases A; D054467:Phospholipases A2; C028213:3-((3-cholamidopropyl)dimethylammonium)-1-propanesulfonate