Quantitative Electroimmunoblotting Study of the Calcium-Activated Neutral Protease in Human Myelin
| Autor: | Claude C.A. Bernard, P.R. Carnegie, N. Kerlero de Rosbo |
|---|---|
| Rok vydání: | 2006 |
| Předmět: |
Multiple Sclerosis
medicine.medical_treatment Endogeny Calcium-Activated Neutral Protease Biochemistry Cellular and Molecular Neuroscience Myelin medicine Humans Egtazic Acid Incubation Edetic Acid Myelin Sheath Mercaptoethanol chemistry.chemical_classification Gel electrophoresis Protease biology Calpain Temperature Brain Myelin Basic Protein Hydrogen-Ion Concentration Myelin basic protein Enzyme medicine.anatomical_structure chemistry Immunologic Techniques biology.protein Calcium Electrophoresis Polyacrylamide Gel |
| Zdroj: | Journal of Neurochemistry. 47:1007-1012 |
| ISSN: | 0022-3042 |
| DOI: | 10.1111/j.1471-4159.1986.tb00713.x |
| Popis: | Degradation of myelin basic protein (MBP) in human man myelin was monitored by electroimmunoblotting. Problems of variation between, as well as within, electroimmunoblots were overcome by the introduction of an internal standard in each sample, thus allowing reproducible quantification of MBP. The Ca2+-dependent protease acting on MBP was active at endogenous levels of Ca2+ (congruent to 300 micrograms/g myelin) and was inhibited in the presence of Ca2+ chelators. Extensive degradation of MBP occurred rapidly in the presence of added Ca2+, reaching a plateau after a 1 h incubation (80-85% degradation). The proteolytic activity was not enhanced in the presence of 2-mercaptoethanol. It was most active at neutral pH and at temperatures approaching physiological conditions. No difference was observed between proteolytic activities of control and multiple sclerotic myelin. It is suggested that fluctuations in the accessibility of free Ca2+ to the protease may lead to the regulation of Ca2+-activated myelinolysis. |
| Databáze: | OpenAIRE |
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