Alteration of the myometrial plasma membrane cholesterol content with beta-cyclodextrin modulates the binding affinity of the oxytocin receptor - PubMed (original) (raw)
. 1995 Oct 24;34(42):13784-93.
doi: 10.1021/bi00042a009.
Affiliations
- PMID: 7577971
- DOI: 10.1021/bi00042a009
Alteration of the myometrial plasma membrane cholesterol content with beta-cyclodextrin modulates the binding affinity of the oxytocin receptor
U Klein et al. Biochemistry. 1995.
Abstract
To investigate the effect of cholesterol on the oxytocin receptor function in myometrial membranes, we developed a new method to alter the membrane cholesterol content. Using a methyl-substituted beta-cyclodextrin, we were able to selectively deplete the myometrial plasma membrane of cholesterol. Vice versa, incubating cholesterol-depleted membranes with a preformed soluble cholesterol-methyl-beta-cyclodextrin complex restored the cholesterol content of the plasma membrane. Binding experiments showed that, with the removal of cholesterol from the membrane, the dissociation constant for [3H]oxytocin is enhanced 87-fold (from Kd = 1.5 nM to Kd = 131 nM), therefore shifting the oxytocin receptor from high to low affinity. Increasing the cholesterol content of the cholesterol-depleted membrane again restored the high-affinity binding (Kd = 1.2 nM). The presence of 0.1 mM GTP gamma S did not significantly change the number of high-affinity binding sites for [3H]oxytocin in native plasma membranes, in membranes depleted of cholesterol, and in plasma membranes with restored cholesterol content. The number of high-affinity binding sites for the oxytocin antagonist [3H]PrOTA was dependent in the same way on the cholesterol content as for [3H]oxytocin. Substitution of the membrane cholesterol with other steroids showed a strong dependence of the oxytocin receptor function on the structure of the cholesterol molecule. The detergent-solubilized oxytocin receptor was not saturable with [3H]oxytocin even at concentrations up to 10(-6) M of radioligand. Addition of the cholesterol-methyl-beta-cyclodextrin complex to the detergent-solubilized oxytocin receptor induced a saturation of the solubilized binding sites (Bmax = 0.98 pmol/mg) for oxytocin (Kd = 16 nM).(ABSTRACT TRUNCATED AT 250 WORDS)
Similar articles
- Conversion of the myometrial oxytocin receptor from low to high affinity state by cholesterol.
Fahrenholz F, Klein U, Gimpl G. Fahrenholz F, et al. Adv Exp Med Biol. 1995;395:311-9. Adv Exp Med Biol. 1995. PMID: 8713981 - Expression of the human oxytocin receptor in baculovirus-infected insect cells: high-affinity binding is induced by a cholesterol-cyclodextrin complex.
Gimpl G, Klein U, Reiländer H, Fahrenholz F. Gimpl G, et al. Biochemistry. 1995 Oct 24;34(42):13794-801. doi: 10.1021/bi00042a010. Biochemistry. 1995. PMID: 7577972 - Reconstitution of the myometrial oxytocin receptor into proteoliposomes. Dependence of oxytocin binding on cholesterol.
Klein U, Fahrenholz F. Klein U, et al. Eur J Biochem. 1994 Mar 1;220(2):559-67. doi: 10.1111/j.1432-1033.1994.tb18656.x. Eur J Biochem. 1994. PMID: 8125115 - Mechanisms regulating oxytocin receptor coupling to phospholipase C in rat and human myometrium.
Sanborn BM, Qian A, Ku CY, Wen Y, Anwer K, Monga M, Singh SP. Sanborn BM, et al. Adv Exp Med Biol. 1995;395:469-79. Adv Exp Med Biol. 1995. PMID: 8713999 Review. - Cholesterol and steroid hormones: modulators of oxytocin receptor function.
Gimpl G, Wiegand V, Burger K, Fahrenholz F. Gimpl G, et al. Prog Brain Res. 2002;139:43-55. doi: 10.1016/s0079-6123(02)39006-x. Prog Brain Res. 2002. PMID: 12436925 Review.
Cited by
- Resveratrol ameliorates high-glucose-induced hyperpermeability mediated by caveolae via VEGF/KDR pathway.
Tian C, Zhang R, Ye X, Zhang C, Jin X, Yamori Y, Hao L, Sun X, Ying C. Tian C, et al. Genes Nutr. 2013 Mar;8(2):231-9. doi: 10.1007/s12263-012-0319-1. Epub 2012 Sep 16. Genes Nutr. 2013. PMID: 22983702 Free PMC article. - Characteristics of pyrene phospholipid/gamma-cyclodextrin complex.
Tanhuanpää K, Cheng KH, Anttonen K, Virtanen JA, Somerharju P. Tanhuanpää K, et al. Biophys J. 2001 Sep;81(3):1501-10. doi: 10.1016/S0006-3495(01)75804-3. Biophys J. 2001. PMID: 11509363 Free PMC article. - Introduction of phospholipids to cultured cells with cyclodextrin.
Kainu V, Hermansson M, Somerharju P. Kainu V, et al. J Lipid Res. 2010 Dec;51(12):3533-41. doi: 10.1194/jlr.D009373. Epub 2010 Sep 29. J Lipid Res. 2010. PMID: 20881052 Free PMC article. - Endosome to Golgi transport of ricin is regulated by cholesterol.
Grimmer S, Iversen TG, van Deurs B, Sandvig K. Grimmer S, et al. Mol Biol Cell. 2000 Dec;11(12):4205-16. doi: 10.1091/mbc.11.12.4205. Mol Biol Cell. 2000. PMID: 11102518 Free PMC article. - Redefining cholesterol's role in the mechanism of the cholesterol-dependent cytolysins.
Giddings KS, Johnson AE, Tweten RK. Giddings KS, et al. Proc Natl Acad Sci U S A. 2003 Sep 30;100(20):11315-20. doi: 10.1073/pnas.2033520100. Epub 2003 Sep 19. Proc Natl Acad Sci U S A. 2003. PMID: 14500900 Free PMC article.
Publication types
MeSH terms
Substances
LinkOut - more resources
Other Literature Sources
Medical