Structural and Biological Stability of the Human Interleukin 10 Homodimer
| Autor: | Eric Huang, Nicholas J. Murgolo, Philip Mui, Rosalinda Syto, William T Windsor, Emory H. Braswell |
|---|---|
| Rok vydání: | 1998 |
| Předmět: |
Interleukin 2
Protein Denaturation Circular dichroism Protein Conformation Stereochemistry Protein subunit Dimer Centrifugation Isopycnic Biochemistry chemistry.chemical_compound Protein structure medicine Humans Receptor Circular Dichroism Biological activity Hydrogen-Ion Concentration Recombinant Proteins Interleukin-10 Interleukin 10 Cross-Linking Reagents chemistry Chromatography Gel Dimerization Ultracentrifugation medicine.drug |
| Zdroj: | Biochemistry. 37:16943-16951 |
| ISSN: | 1520-4995 0006-2960 |
| Popis: | Human interleukin 10 (huIL-10) is a cytokine that regulates the synthesis of type 1 helper T cell derived cytokines such as gamma-interferon, interleukin 2, and tumor necrosis factor alpha. The potential immunosuppressive activities of huIL-10 suggest that this protein may be clinically useful for treating autoimmune diseases. Due to the potential clinical value of this cytokine, physicochemical studies have been performed regarding its association state and biological/structural stability. These studies include performing size-exclusion chromatography, chemical cross-linking, equilibrium ultracentrifugation, and circular dichroism spectroscopy. The results indicate huIL-10 is predominantly a noncovalent homodimer at neutral pH and 4 degreesC for concentrations greater than 0.003 mg/mL (0.08 microM dimer). An apparent pKa value of approximately 4.8 was calculated for both the pH-dependent subunit dissociation and pH-induced loss in MC/9 biological activity. A temperature analysis revealed a linear relationship between the percent dimer and relative MC/9 activity, thus, these results and the pH-dependent activity results suggest that the huIL-10 dimer is the active species. The GndHCl-induced unfolding of rhuIL-10, monitored by far-UV circular dichroism, revealed a unique biphasic unfolding process which contained both a subunit dissociation process ( |
| Databáze: | OpenAIRE |
| Externí odkaz: |
|