Search

Showing total 4 results
Start Over Topic intermediate filament proteins Journal journal of neuroimmunology
4 results

1. Varying degrees of phosphorylation determine microheterogeneity of the heavy neurofilament polypeptide (Nf-H).

Author

Goldstein ME, Sternberger LA, and Sternberger NH

Subjects
Antibodies, Antibodies, Monoclonal, Collodion, Electrophoresis, Polyacrylamide Gel, Epitopes analysis, Neurofilament Proteins, Paper, Phosphorylation, Protein Processing, Post-Translational, Staining and Labeling, Intermediate Filament Proteins metabolism
Abstract

Two-dimensional immunoblots revealed a spectrum of 200 kDa neurofilament polypeptides (Nf-H) of apparent molecular weights ranging from 200 to 170 kDa. The entire spectrum was stained immunocytochemically by three monoclonal antibodies specific for nonphosphorylated neurofilaments, while more restricted staining was revealed by four monoclonal antibodies specific for phosphorylated neurofilament epitopes. Treatment with increasing amounts of phosphatase suggested the existence of various forms of partially phosphorylated neurofilaments that possess phosphoepitopes that differ in their ease of dephosphorylation. Immunoprecipitation in low detergent concentration confirmed the existence of microheterogeneous forms of Nf-H that differed in extent of phosphorylation or in distribution of phosphorylated sites.

Published
1987
Full Text
View/download PDF

2. Phosphorylation protects neurofilaments against proteolysis.

Author

Goldstein ME, Sternberger NH, and Sternberger LA

Subjects
Chromatography, Affinity, Chymotrypsin metabolism, Collodion, Cytoskeleton metabolism, Drug Stability, Electrophoresis, Polyacrylamide Gel, Neurofilament Proteins, Paper, Phosphoric Monoester Hydrolases metabolism, Phosphorylation, Intermediate Filament Proteins metabolism
Abstract

During incubation with phosphatase, the 200 kDa neurofilament protein in cytoskeletal preparations is degraded extensively. Degradation, which is divalent cation-independent, does not occur when inhibitors of phosphatase are added. The 160 kDa chymotryptic fragment of neurofilaments or affinity-purified 200 kDa protein are not degraded by phosphatase. The results suggest that phosphorylated neurofilaments are protected against proteolysis, and dephosphorylated neurofilaments are degraded by a calcium-independent, endogenous proteinase which is associated with assembled neurofilaments or with other cytoskeletal components, and not with the phosphatase used.

Published
1987
Full Text
View/download PDF

3. Varying degrees of phosphorylation determine microheterogeneity of the heavy neurofilament polypeptide (Nf-H)

Author

Margi E. Goldstein, Nancy H. Sternberger, and Ludwig A. Sternberger

Subjects
Paper, Neurofilament, Immunoprecipitation, medicine.drug_class, Immunology, Phosphatase, Monoclonal antibody, Antibodies, Epitope, Dephosphorylation, Epitopes, Intermediate Filament Proteins, Neurofilament Proteins, medicine, Immunology and Allergy, Phosphorylation, Staining and Labeling, Molecular mass, Chemistry, Antibodies, Monoclonal, Collodion, Molecular biology, Neurology, Biochemistry, Electrophoresis, Polyacrylamide Gel, Neurology (clinical), Protein Processing, Post-Translational
Abstract

Two-dimensional immunoblots revealed a spectrum of 200 kDa neurofilament polypeptides (Nf-H) of apparent molecular weights ranging from 200 to 170 kDa. The entire spectrum was stained immunocytochemically by three monoclonal antibodies specific for nonphosphorylated neurofilaments, while more restricted staining was revealed by four monoclonal antibodies specific for phosphorylated neurofilament epitopes. Treatment with increasing amounts of phosphatase suggested the existence of various forms of partially phosphorylated neurofilaments that posses phosphoepitopes that differ in their ease of dephosphorylation. Immunoprecipitation in low detergent concentration confirmed the existence of microheterogeneous forms of Nf-H that differed in extent of phosphorylation or in distribution of phosphorylated sites.

Published
1987

4. Phosphorylation protects neurofilaments against proteolysis

Author

Ludwig A. Sternberger, Margi E. Goldstein, and Nancy H. Sternberger

Subjects
Paper, Neurofilament, Proteolysis, Immunology, Immunocytochemistry, Phosphatase, Endogeny, Biology, Chromatography, Affinity, Divalent, Drug Stability, Intermediate Filament Proteins, Neurofilament Proteins, medicine, Chymotrypsin, Immunology and Allergy, Phosphorylation, Cytoskeleton, chemistry.chemical_classification, medicine.diagnostic_test, Collodion, Phosphoric Monoester Hydrolases, Neurology, Biochemistry, chemistry, Electrophoresis, Polyacrylamide Gel, Neurology (clinical)
Abstract

During incubation with phosphatase, the 200 kDa neurofilament protein in cytoskeletal preparations is degraded extensively. Degradation, which is divalent cation-independent, does not occur when inhibitors of phosphatase are added. The 160 kDa chymotryptic fragment of neurofilaments or affinity-purified 200 kDa protein are not degraded by phosphatase. The results suggest that (1) phosphorylated neurofilaments are protected against proteolysis, and (2) dephosphorylated neurofilaments are degraded by a calcium-independent, endogenous proteinase which is associated with assembled neurofilaments or with other cytoskeletal components, and not with the phosphatase used.

Published
1987