Preferential action of a brain detyrosinolating carboxypeptidase on polymerized tubulin.

N. Kumar, M. Flavin

Research output: Contribution to journalArticle

Abstract

A carboxypeptidase purified from brain catalyzes the release of COOH-terminal tyrosine without further digesting tubulin. It is distinct from previously described carboxypeptidases, and appears to have specificity for tubulin as it is not inhibited by peptides and proteins with COOH-terminal tyrosine, and because, unlike carboxypeptidase A (which by removing tyrosine from aldolase causes its inactivation), this enzyme does not decrease aldolase activity. The enzyme detyrosinolates both self-assembly-competent (cycle-purified) and -incompetent (phosphocellulose-purified) tubulin. However, under assembly conditions the rate was 2-3-fold higher for competent tubulin. Preincubation of assembly-competent tubulin with podophyllotoxin or colchicine resulted in a parallel concentration-dependent inhibition of tubulin polymerization and detyrosinolation. Similarly, when incompetent tubulin was induced to polymerize by preincubation with purified microtubule-associated protein 2 (an assembly-promoting protein) or taxol, the initial rate of its detyrosinolation increased 3-5-fold, and this increase was blocked if podophyllotoxin was also added along with microtubule-associated protein 2 or taxol during the preincubation. Oligomers induced by adding vinblastine to incompetent tubulin were also detyrosinolated more rapidly, and the stimulation was abolished by maytansine, which has been shown to disperse the vinblastine-induced oligomers. When polymerized and subunit fractions were separated after a steady state mixture had been partially digested with the carboxypeptidase, the former was found to have lost 2-3 times more COOH-terminal tyrosine. Although both polymer and monomer can be detyrosinolated by the enzyme, polymeric and oligomeric forms are the preferred substrates. Carboxypeptidase appeared to release tyrosine at the same rate from populations of short and long microtubules.

Original languageEnglish (US)
Pages (from-to)7678-7686
Number of pages9
JournalJournal of Biological Chemistry
Volume256
Issue number14
StatePublished - Jul 25 1981
Externally publishedYes

Fingerprint

Carboxypeptidases
Tubulin
Brain
Tyrosine
Podophyllotoxin
Fructose-Bisphosphate Aldolase
Microtubule-Associated Proteins
Vinblastine
Paclitaxel
Oligomers
Maytansine
Enzymes
Carboxypeptidases A
Colchicine
Microtubules
Polymerization
Self assembly
Polymers
Proteins
Monomers

ASJC Scopus subject areas

  • Biochemistry

Cite this

Preferential action of a brain detyrosinolating carboxypeptidase on polymerized tubulin. / Kumar, N.; Flavin, M.

In: Journal of Biological Chemistry, Vol. 256, No. 14, 25.07.1981, p. 7678-7686.

Research output: Contribution to journalArticle

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