Osmolytes improve the reconstitution of luteinizing hormone/ human chorionic gonadotropin receptors into proteoliposomes

Kolena, J; Matejčíková, K.; šrenkelová, G

doi:10.1016/0303-7207(92)90160-8

Cited by 8 publications

(10 citation statements)

References 30 publications

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“…These substances stabilized proteins against thermal denaturation (Arakawa and Timasheff, 1985). Heat inactivation of LH/hCG-binding sites demonstrated a significant stabilization of the receptor structure in the presence of osmolytes, follicular fluid , serum and albumin (Kolena et al, 1992;Kolena et al, 1999). The present paper indicates that this stabilizing effect on the LH/hCG receptor may be caused by elimination of negative membrane charge yielded by free fatty acids.…”

Section: Introductionmentioning

confidence: 56%

“…The stability of proteins in their native conformation has become a topic increasing attention over the last several years. Previous studies demonstrated that glycerol and other osmolytes had beneficial effects on preservation of the activity of LH/hCG receptors (Metsikko et al, 1990;Kolena et al, 1992). These substances stabilized proteins against thermal denaturation (Arakawa and Timasheff, 1985).…”

Section: Introductionmentioning

confidence: 99%

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Thermal destabilization of ovarian LH/hCG receptors by negatively charged lipids

Kolena

Scsuková²,

Ježová³

2002

Exp Clin Endocrinol Diabetes

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The stabilizing effect of BSA on the rat ovarian LH/hCG receptor was analyzed by thermal perturbation technique. Thermal destabilization of the receptor with arachidonic acid along with digestion of membrane with phospholipase A2 and reversal of these effects when BSA was used as fatty acids scavenger, may indicate that free fatty acids are responsible for instability of the LH/hCG receptor. This destabilizing effect may be caused by the presence of a net negative surface charge provided by fatty acids. This presumption was corroborated by the reconstitution of delipidated LH/hCG receptor into proteoliposomes. Delipidated receptor lost to a great extent its binding activity and thermal stability. The receptor was fully reactivated by the reconstitution into proteoliposomes with neutral phosphatidylcholine but not with negatively charged phosphatidylserine and phosphatidylglycerol. Thermal inactivation of the LH/hCG receptor by delipidation was entirely inverted by treatment with phosphatidylcholine but the presence of negatively charged phospholipids did not change the heat inactivation profile of hCG-binding sites.

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Section: Introductionmentioning

confidence: 56%

Section: Introductionmentioning

confidence: 99%

Thermal destabilization of ovarian LH/hCG receptors by negatively charged lipids

Kolena

Scsuková²,

Ježová³

2002

Exp Clin Endocrinol Diabetes

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“…1 -I NaCI, pH 7.4) containing 20% sucrose were centrifuged at 1,000 x g for 15 rain and the supernatant was further centrifuged at 20,000 x g for 30 min. Plasma membranes were prepared on the sucrose gradient [8,13]. Reconstitution of the ovarian LH/hCG receptor into proteoliposomes after removal of sodium cholate detergent by absorption on Bin-Beads SM-2 was described previously [13,14].…”

Section: Methodsmentioning

confidence: 99%

“…Plasma membranes were prepared on the sucrose gradient [8,13]. Reconstitution of the ovarian LH/hCG receptor into proteoliposomes after removal of sodium cholate detergent by absorption on Bin-Beads SM-2 was described previously [13,14]. Isolated luteal cells were prepared by enzymatic dispersion of luteinized ovaries with collagenase [15].…”

Section: Methodsmentioning

confidence: 99%

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Early hCG‐induced desensitization of rat ovarian LH/hCG receptors is associated with altered physical state of membranes

1994

FEBS Letters

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Injection of pseudopregnant rats with pharmacological doses of hCG produced desensitization of adenylylcyclase and steroidogenic systems in ovarian membranes and luteal cells. Membrane lipid rigidity, as determined by fluorescence polarization of DPH, decreased as early as 0.5 h after injection of hCG. Desensitization also modified the differential scanning calorimetric profile characteristic of control membranes. The accessibility of ovarian LH/hCG receptors was unchanged. The results indicate that the hCG-induced decrease of membrane lipid rigidity is preceded by the process of desensitization of the rat ovary.

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Paramagnetic Centers in the Organism of Volunteers and Patients in Various Pathological States

2002

Electron Paramagnetic Resonance in Biochemistry and Medicine

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Osmolytes improve the reconstitution of luteinizing hormone/ human chorionic gonadotropin receptors into proteoliposomes

Cited by 8 publications

References 30 publications

Thermal destabilization of ovarian LH/hCG receptors by negatively charged lipids

Thermal destabilization of ovarian LH/hCG receptors by negatively charged lipids

Early hCG‐induced desensitization of rat ovarian LH/hCG receptors is associated with altered physical state of membranes

Paramagnetic Centers in the Organism of Volunteers and Patients in Various Pathological States

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