Solubilization of [35S]lanthionine ketimine binding sites from bovine brain.
Lanthionine ketimine (LK) binding sites were solubilized from bovine brain membranes using 3-[(3-cholamidopropyl)dimethylammonio]-1-propanesulfonate (CHAPS) and Triton X-100. 10 mM CHAPS in 0.5 M potassium phosphate, pH 7.0, containing 20% glycerol was selected to solubilize LK binding entities. Some properties of CHAPS-solubilized LK binding sites have been studied. The CHAPS-solubilized preparation appeared to contain a homogenous population of binding sites for [35S]LK. Binding properties indicated that the solubilized binding sites were similar to the membrane-bound sites. [35S]LK specific binding was inhibited by other structurally related ketimines obtaining a similar rank order of inhibition for the soluble and the membrane-bound preparations. The successful solubilization of [35S]LK binding sites is a useful starting point for the purification of this binding protein.
Fontana
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M
Dipartimento di Scienze Biochimiche, Università di Roma, Italia.
Costa
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M
Duprè
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S
D000603:Amino Acids, Sulfur / Q000378:metabolism*; D000818:Animals; D001665:Binding Sites / Q000187:drug effects / Q000502:physiology; D001667:Binding, Competitive / Q000187:drug effects; D001923:Brain Chemistry / Q000187:drug effects / Q000502:physiology*; D002417:Cattle; D002793:Cholic Acids; D066298:In Vitro Techniques; D007700:Kinetics; D008566:Membranes / Q000737:chemistry; D017830:Octoxynol; D011485:Protein Binding; D011869:Radioligand Assay; D012995:Solubility; D013438:Sulfhydryl Compounds / Q000494:pharmacology; D013462:Sulfur Radioisotopes
D000603:Amino Acids, Sulfur; D002793:Cholic Acids; D013438:Sulfhydryl Compounds; D013462:Sulfur Radioisotopes; C035499:lanthionine ketimine; D017830:Octoxynol; C028213:3-((3-cholamidopropyl)dimethylammonium)-1-propanesulfonate