ADP-ribosylation of microtubule proteins as catalyzed by cholera toxin
| Autor: | U.Z. Littauer, Yoram Salomon, E. Ezra, A. Zutra, Y. Amir-Zaltsman, T. Scherson |
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| Jazyk: | angličtina |
| Rok vydání: | 1982 |
| Předmět: |
Male
Cholera Toxin Microtubule-associated protein medicine.disease_cause General Biochemistry Genetics and Molecular Biology chemistry.chemical_compound Tubulin Endopeptidases medicine Animals Sodium dodecyl sulfate Molecular Biology Polyacrylamide gel electrophoresis Brain Chemistry Adenosine Diphosphate Ribose General Immunology and Microbiology biology Isoelectric focusing Nucleoside Diphosphate Sugars General Neuroscience Cholera toxin Serine Endopeptidases Proteins NAD Molecular biology Rats Kinetics Isoelectric point Biochemistry chemistry ADP-ribosylation biology.protein Electrophoresis Polyacrylamide Gel Microtubule-Associated Proteins Research Article |
| Popis: | Incubation of purified rat brain tubulin with cholera toxin and radiolabeled [32P] or [8-3H]NAD results in the labeling of both alpha and beta subunits as revealed on sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE). Treatment of these protein bands with snake venom phosphodiesterase resulted in quantitative release of labeled 5'-AMP, respectively labeled with the corresponding isotope. Two-dimensional separation by isoelectric focusing and SDS-PAGE of labeled and native tubulin revealed that labeling occurs at least in four different isotubulins. The isoelectric point of the labeled isotubulins was slightly lower than that of native purified tubulin. This shift in mobility is probably due to additional negative charges involved with the incorporation of ADP-ribosyl residues into the tubulin subunits. SDS-PAGE of peptides derived from [32P]ADP-ribosylated alpha and beta tubulin subunits by Staphylococcus aureus protease cleavage showed a peptide pattern identical with that of native tubulin. Microtubule-associated proteins (MAP1 and MAP2) of high molecular weight were also shown to undergo ADP-ribosylation. Incubation of permeated rat neuroblastoma cells in the presence of [32P]NAD and cholera toxin results in the labeling of only a few cell proteins of which tubulin is one of the major substrates. |
| Databáze: | OpenAIRE |
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