[Purification to apparent homogeneity of the somatostatin receptor of the human cell line HGT-1 of gastric origin].
This communication reports the isolation and the purification of the gastric somatostatin receptor from the human cell line HGT-1. The receptor has been extracted from the cell membrane by Triton X 100, and a monoclonal antibody to this was prepared. A series of affinity chromatographies (Sepharose-antibody and Sepharose-somatostatin-14) and a final purification by steric exclusion on high performance liquid chromatography columns (HPLC) allowed us to obtain a fraction enriched 20,000 fold in 125I-Tyrll-somatostatin-14 specific binding (apparent dissociation constant: 7.6 x 10(-8) M). This fraction corresponded to a molecular mass of about 90 kDa (in presence of detergent) and to a maximal binding capacity of more than 10,000 pmol/protein. It therefore has a theoretical homogeneity close to 100%.
Reyl-Desmars
F
F
Unité de Recherche de Gastroentérologie, I.N.S.E.R.M. U10, Hôpital Bichat, Paris.
Le Roux
S
S
Linard
C
C
Benkouka
F
F
Lewin
M J
MJ
D002462:Cell Membrane / Q000032:analysis; D002846:Chromatography, Affinity; D002851:Chromatography, High Pressure Liquid; D006801:Humans; D015151:Immunoblotting; D007124:Immunoenzyme Techniques; D008970:Molecular Weight; D017981:Receptors, Neurotransmitter / Q000302:isolation & purification* / Q000378:metabolism; D017481:Receptors, Somatostatin; D013004:Somatostatin / Q000031:analogs & derivatives / Q000378:metabolism; D013274:Stomach Neoplasms / Q000032:analysis*; D014407:Tumor Cells, Cultured
D017981:Receptors, Neurotransmitter; D017481:Receptors, Somatostatin; D013004:Somatostatin; C012088:somatostatin, N-Tyr(1)-
Purification jusqu'à apparente homogénéité du récepteur de la somatostatine de la lignée cellulaire humaine HGT-1 d'origine gastrique.