Solubilization of the ecdysone binding protein from anterior silk gland cell membranes of the silkworm, Bombyx mori.
We previously provided preliminary evidence for the presence of a putative membrane ecdysone receptor (mEcR) anchored in the plasma membranes of anterior silk glands (ASGs) in Bombyx mori. This receptor may act in concert with the conventional EcR in 20E-dependent programmed cell death of these glands. We report here, for the first time, the solubilization of mEcR from ASG membranes using the zwitterionic detergent CHAPS in the presence of NaCl. Our results show by ligand binding assay that mEcR solubilized this way is functionally active and retains 75% of its native binding activity. We also defined experimental conditions that yielded protein/detergent complexes with partial binding activity, which makes it possible to purify the membrane-bound ecdysone binding protein.
Elmogy
Mohamed
M
Division of Life Sciences, Graduate School of Science and Technology, Kanazawa University, Kakumamachi, Japan.
Iwami
Masafumi
M
Sakurai
Sho
S
D000818:Animals; D012831:Bombyx / Q000502:physiology*; D002793:Cholic Acids; D003902:Detergents; D008565:Membrane Proteins / Q000302:isolation & purification* / Q000378:metabolism; D011987:Receptors, Steroid / Q000302:isolation & purification* / Q000378:metabolism; D012965:Sodium Chloride / Q000737:chemistry; D012995:Solubility; D013347:Subcellular Fractions / Q000737:chemistry; D014316:Tritium
D002793:Cholic Acids; D003902:Detergents; D008565:Membrane Proteins; D011987:Receptors, Steroid; C055469:ecdysone receptor; D014316:Tritium; D012965:Sodium Chloride; C028213:3-((3-cholamidopropyl)dimethylammonium)-1-propanesulfonate