| Autor: |
Sperbeck SJ; Department of Obstetrics and Gynecology, University of Cincinnati College of Medicine, OH 45267-0526., Song DY, LaBarbera AR |
| Jazyk: |
angličtina |
| Zdroj: |
Journal of receptor research [J Recept Res] 1993; Vol. 13 (6), pp. 925-42. |
| DOI: |
10.3109/10799899309073701 |
| Abstrakt: |
Follicle-stimulating hormone (FSH)-receptors were solubilized from immature porcine ovarian granulosa cells with retention of high affinity 125I-porcine FSH-binding activity. The optimal concentration of Triton X-100 for solubilization was 0.5% (w/v), and the optimal cellular protein concentration 25 mg/ml. Glycerol (30%) increased recovery of solubilized receptor. 125I-pFSH-binding affinity ranged from 4 x 10(10) M-1 to 8 x 10(10) M-1 in either the absence or presence of glycerol. 125I-pFSH-binding capacity was 5 fmol/mg protein in the absence of glycerol and 58 fmol/mg protein in the presence of glycerol as determined by equilibrium saturation binding analysis. By gel permeation chromatography, the apparent size of the 125I-pFSH-receptor complex was 462 kDa in the absence of glycerol and 762 kDa in the presence of glycerol. Ligand blotting of solubilized receptor yielded a single species with an apparent molecular weight of 200 kDa under nonreducing conditions and a single species with an apparent molecular weight of 60 kDa under reducing conditions. These studies indicated that high affinity FSH-binding activity can be solubilized from membranes of immature porcine granulosa. |
| Databáze: |
MEDLINE |
| Externí odkaz: |
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