Isolation and identification of proteolipid proteins injimpy mouse brain
| Autor: | F. G. Mastronardi, Mario A. Moscarello, A. M. Fannon |
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| Rok vydání: | 1994 |
| Předmět: |
Ratón
Proteolipids Molecular Sequence Data Biology medicine.disease_cause Biochemistry Mice Cellular and Molecular Neuroscience Myelin immune system diseases Complementary DNA medicine Animals Amino Acid Sequence Amino Acids Myelin Proteolipid Protein Brain Chemistry chemistry.chemical_classification Mutation Mice Jimpy Myelin Basic Protein General Medicine nervous system diseases Amino acid Blot medicine.anatomical_structure chemistry biology.protein lipids (amino acids peptides and proteins) Antibody Myelin Proteins |
| Zdroj: | Neurochemical Research. 19:1005-1012 |
| ISSN: | 1573-6903 0364-3190 |
| Popis: | Proteolipids were isolated from 20 day old normal and jimpy mouse brain by extraction into chloroform-methanol (2:1, w/v), delipidated by size-exclusion HPLC, and analyzed by SDS-PAGE, Western blots, amino acid analyses, and N-terminal sequencing. SDS-PAGE showed that a major proteolipid from jimpy mouse brain had an apparent molecular weight of approximately 23 kDa, intermediate to that of PLP and DM-20 from normal mouse brain. Western blots with 3 different antibodies which recognize residues 200-224, 116-150, and 270-276 respectively recognized immunoreactive material in normal and jimpy PLP. Since antibody reactive with 270-276 did not recognize jimpy PLP, an altered C-terminus of the jimpy protein is suggested. These results demonstrated that a PLP can be partially purified from jimpy mouse brain. Amino acid analyses failed to show the predicted increase in cysteinyl residues (predicted from cDNA) in jimpy PLP. However, when jimpy brain proteolipids were subjected to N-terminal sequencing, Gly, Leu, Leu, Gly the first four amino acids of PLP were detected. Thus, the partial purification of a proteolipid from jimpy mouse brain, whose characteristics (apparent molecular weight, immunoreactivity, N-terminal sequence and relative net charge) strongly suggested that PLP of altered size is present in jimpy mouse brain. |
| Databáze: | OpenAIRE |
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