[Solubilization, purification and molecular characterization of H2 histamine receptor from human tumoral gastric cells HGT-1].
This communication reports the solubilization, the purification and the molecular characterization of the H2-histamine receptor from the cell line HGT-1 derived from a human gastric cancer. The receptor has been solubilized by Triton X100 and purified by gel filtration onto Sephacryl, affinity-chromatography (Sepharose-famotidine) and high performance liquid chromatography (HPLC). The purified receptor specifically bound the H2 selective ligand 3H-methyltiotidine with a kD of 160 nM (vs 50 nM for the intact HGT-1 cell) and a maximal binding capacity of 14,000 pmol/mg protein which represents a 12,170-fold enrichment and a degree of purity of 98%. It is a glycoprotein of 70 kDa molecular mass containing N-acetylglucosamine residues.
Reyl-Desmars
F
F
Unité de Recherches de Gastroentérologie, I.N.S.E.R.M. U n. 10, Hôpital Bichat, Paris.
Cherifi
Y
Y
Le Romancer
M
M
Pigeon
C
C
Le Roux
S
S
Lewin
M J
MJ
D002461:Cell Line, Transformed; D002471:Cell Transformation, Neoplastic / Q000737:chemistry*; D002846:Chromatography, Affinity; D002850:Chromatography, Gel; D002851:Chromatography, High Pressure Liquid; D006801:Humans; D011092:Polyethylene Glycols; D011970:Receptors, Histamine H2 / Q000302:isolation & purification*; D012995:Solubility; D013274:Stomach Neoplasms / Q000473:pathology*
D011970:Receptors, Histamine H2; D011092:Polyethylene Glycols
Solubilisation, purification et caractérisation moléculaire du récepteur histaminique H2 à partir des cellules tumorales gastriques humaines HGT-1.