Tubulin carboxypeptidase/microtubules association can be detected in the distal region of neural processes

The association of tubulin carboxypeptidase with microtubules has been demonstrated in crude brain extracts and in living non-nervous cells. Here, we studied this phenomenon in cultured brain cells. To determine the association of the enzyme with neural microtubules we isolated the cytoskeletons (de...

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Bibliographic Details
Published in:Neurochemical research 2000, Vol.25 (1), p.27-36
Main Authors: CONTIN, M. A, ARCE, C. A
Format: Article
Language:English
Subjects:
Animals
Biochemistry and metabolism
Biological and medical sciences
Brain - enzymology
Brain - ultrastructure
Carboxypeptidases - metabolism
Cells, Cultured
Central nervous system
Chick Embryo
Fluorescent Antibody Technique
Fundamental and applied biological sciences. Psychology
Glutamine - analysis
Glutamine - metabolism
Microtubules - enzymology
Microtubules - metabolism
Paclitaxel - pharmacology
Time Factors
Tubulin - analysis
Tubulin - metabolism
tubulin carboxypeptidase
Tyrosine - analysis
Tyrosine - metabolism
Vertebrates: nervous system and sense organs
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Summary:The association of tubulin carboxypeptidase with microtubules has been demonstrated in crude brain extracts and in living non-nervous cells. Here, we studied this phenomenon in cultured brain cells. To determine the association of the enzyme with neural microtubules we isolated the cytoskeletons (detergent-extraction under microtubule-stabilizing conditions) and measured the content of Tyr, Glu, and delta2 tubulin as a function of the in vitro incubation time of the cytoskeletons. The carboxypeptidase was found associated with microtubules in 2 days-cultured cells but not in 7 days-cultured cells. Quantitative analysis of digitized images after immunofluorescent staining revealed that detyrosination during the incubation of the cytoskeletons occurred preferentially in the distal regions of the neural processes. Prolonged taxol-treatment of the cells promoted higher detyrosination but Tyr tubulin was not depleted suggesting the existence of a subset of microtubules that has not associated carboxypeptidase and therefore cannot be detyrosinated even after prolonged taxol-treatment. This hypothesis was supported, although not conclusively, by additional experiments.
ISSN:0364-3190
1573-6903
DOI:10.1023/A:1007579113813