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Covalent Cross-Linking of Radiolabeled Human Chorionic Gonadotropin to Rat Ovarian Luteinizing Hormone Receptor with Glutaraldehyde
| Content Provider | Scilit |
|---|---|
| Author | Petäjä, Ulla Kellokumpu, Sakari Keinänen, Kari Metsikkö, Kalervo Rajaniemi, Hannu |
| Copyright Year | 1987 |
| Description | Crude plasma membranes of pseudopregnant rat ovaries were incubated with 125I-labeled human chorionic gonadotropin (125I-hCG) and the formed luteinizing hormone (LH)/hCG receptor-125I-hCG complex was solubilized, partially purified by Sepharose 6B gel filtration, cross-linked with glutaraldehyde and subjected to sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and fluorography. An apparent molecular weight (mol wt) of 130,000 was obtained for the non-reduced complex. A similar-sized complex was observed, when 3H-hCG (β-subunit labeled) instead of 125I-hCG (α-subunit labeled) was used, indicating that the complex contains intact hCG. Upon reduction of the cross-linked receptor-125I-hCG complex, a 105,000 mol wt complex in addition to the 130,000 one was observed. No smaller complexes appeared, however, upon reduction of the receptor-3H-hCG complex, suggesting that the α-subunit of hCG predominantly interacts with the receptor. The cross-linking efficiency was dependent on protein concentration, glutaraldehyde concentration, pH, reaction time and temperature. Under optimal conditions (2 mM glutaraldehyde, pH 7.0-8.0, 60 min, 20°C) no nonspecific complexes appeared and the efficiency of cross-linking of the partially purified detergent-solubilized receptor-125I-hCG complex was approximately 30%. Glutaraldehyde thus provides a rapid and efficient cross-linking reagent to covalently attach 125I-hCG to its receptor and is likely to be highly applicaple to other receptor-ligand systems as well. |
| Related Links | http://www.tandfonline.com/doi/pdf/10.3109/10799898709054563 |
| Ending Page | 827 |
| Page Count | 19 |
| Starting Page | 809 |
| ISSN | 01975110 |
| DOI | 10.3109/10799898709054563 |
| Journal | Journal of receptor research |
| Issue Number | 6 |
| Volume Number | 7 |
| Language | English |
| Publisher | Informa UK Limited |
| Publisher Date | 1987-01-01 |
| Access Restriction | Open |
| Subject Keyword | Journal: Journal of Receptor Research Biochemistry and Molecular Biology Analytical Chemistry Luteinizing Hormone Receptor |
| Content Type | Text |
| Resource Type | Article |
| Subject | Pharmacology |
