5-HT1A and histamine H1 receptors in HeLa cells stimulate phosphoinositide hydrolysis and phosphate uptake via distinct G protein pools. J Biol Chem 1991 Jan 05;266(1):372-9
Date
01/05/1991Pubmed ID
1845968Scopus ID
2-s2.0-0025924749 (requires institutional sign-in at Scopus site) 65 CitationsAbstract
Regulation of phosphate uptake was studied in a HeLa cell line after transfection with DNA encoding the human 5-HT1A receptor. In these cells, 5-HT stimulates sodium-dependent phosphate uptake via protein kinase C activation. Endogenous histamine H1 receptors (739 +/- 20 fmol/mg protein) were identified with [3H]pyrilamine. Histamine (i) stimulated phosphoinositide hydrolysis (EC50 = 8.6 +/- 4.1 microM), (ii) activated protein kinase C (2.4-fold increase in activity), and (iii) increased phosphate uptake (EC50 = 3.2 +/- 1.8 microM) by increasing maximal transport (Vmax(basal) = 6.2 +/- 0.3 versus Vmax(histamine) = 9.1 +/- 0.4) without changing the affinity of the transport process for phosphate. Prolonged treatment with 16 microM phorbol 12-myristate 13-acetate completely blocked protein kinase C activation and markedly attenuated the stimulation of phosphate uptake induced by histamine, establishing that 5-HT and histamine stimulate phosphate uptake through the common pathway of protein kinase C activation. The linkages of the histamine H1 and 5-HT1A receptors to G protein pools were assessed in two ways. (i) The stimulation of phosphoinositide hydrolysis, protein kinase C activity, and phosphate uptake associated with histamine were insensitive to pertussis toxin, whereas those associated with 5-HT were very sensitive to pertussis toxin. (ii) The stimulation of phosphoinositide hydrolysis, protein kinase C activity, and phosphate uptake induced by histamine and 5-HT were additive. These findings suggest that distinct receptor types can stimulate phosphoinositide hydrolysis, protein kinase C, and phosphate uptake in an additive fashion through distinct pools of G proteins in a single cell type.
Author List
Raymond JR, Albers FJ, Middleton JP, Lefkowitz RJ, Caron MG, Obeid LM, Dennis VWAuthor
John R. Raymond MD President, CEO, Professor in the President department at Medical College of WisconsinMESH terms used to index this publication - Major topics in bold
Biological TransportBlotting, Northern
Down-Regulation
GTP-Binding Proteins
HeLa Cells
Histamine
Humans
Hydrolysis
Kinetics
Pertussis Toxin
Phorbol 12,13-Dibutyrate
Phosphates
Phosphatidylinositols
Phosphorylation
Protein Kinase C
Pyrilamine
RNA, Messenger
Receptors, Histamine H1
Receptors, Serotonin
Serotonin
Tetradecanoylphorbol Acetate
Transfection
Virulence Factors, Bordetella