Purification to homogeneity and amino acid sequence analysis of a receptor protein for interleukin 1
| Autor: | Kathryn P. Parker, Patricia L. Kilian, Alvin S. Stern, Yu-Ching E. Pan, Richard Anthony Chizzonite, Jeffrey D. Hulmes, Raina S. Hellmann, Dale Mueller |
|---|---|
| Rok vydání: | 1989 |
| Předmět: |
Thymoma
Receptors Antigen T-Cell Biophysics Biology Biochemistry Chromatography Affinity Cell Line Mice Affinity chromatography Complementary DNA Animals Amino Acids Receptors Immunologic Receptor Molecular Biology Peptide sequence Gel electrophoresis Molecular mass Receptors Interleukin-1 Thymus Neoplasms Ligand (biochemistry) Molecular biology Molecular Weight Dissociation constant Kinetics Chromatography Gel Interleukin-1 |
| Zdroj: | Archives of Biochemistry and Biophysics. 274:26-36 |
| ISSN: | 0003-9861 |
| Popis: | The interleukin 1 (IL-1) receptor from mouse EL-4 thymoma cells was purified to homogeneity by a method which utilized ligand affinity chromatography and classical chromatographic techniques. After solubilization of the receptor from intact cells with the zwitterionic detergent 3-[(3-cholamidopropyl)dimethylammonio]-1-propanesulfonate, the IL-1 binding activity was purified greater than 23,000-fold. Analysis of the purified protein by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, immunoblot, and ligand blot demonstrated that a single protein of molecular mass of approximately 80 kDa is the IL-1 binding polypeptide. The purified protein bound IL-1 with a dissociation constant of approximately 1.1 X 10(-10) M, which is indistinguishable from the affinity of the cell-bound receptor. The amino acid composition of this protein is strikingly similar to the composition deduced from the sequence of a cDNA coding for an IL-1 receptor from EL-4 cells. Protein sequence analysis of Staphylococcus aureus V-8 protease-derived peptides yields data consistent with the sequence proposed from cloned cDNA. These studies have demonstrated that the high affinity IL-1 receptor on EL-4 cells is the 80-kDa protein. |
| Databáze: | OpenAIRE |
| Externí odkaz: |
|