Microtubule dysfunction by posttranslational nitrotyrosination of α- tubulin

A nitric oxide-dependent mechanism of cellular injury

J. P. Eiserich, A. G. Estevez, T. V. Bamberg, Zu Ye Yao Zu Ye, P. H. Chumley, J. S. Beckman, B. A. Freeman

Research output: Contribution to journalArticle

224 Citations (Scopus)

Abstract

NO2Tyr (3-Nitrotyrosine) is a modified amino acid that is formed by nitric oxide-derived species and has been implicated in the pathology of diverse human diseases. Nitration of active-site tyrosine residues is known to compromise protein structure and function. Although free NO2Tyr is produced in abundant concentrations under pathological conditions, its capacity to alter protein structure and function at the translational or posttranslational level is unknown. Here, we report that free NO2Tyr is transported into mammalian cells and selectively incorporated into the extreme carboxyl terminus of α-tubulin via a posttranslational mechanism catalyzed by the enzyme tubulin-tyrosine ligase. In contrast to the enzymatically regulated carboxyl-terminal tyrosination/detyrosination cycle of α-tubulin, incorporation of NO2Tyr shows apparent irreversibility. Nitrotyrosination of α-tubulin induces alterations in cell morphology, changes in microtubule organization, loss of epithelial-barrier function, and intracellular redistribution of the motor protein cytoplasmic dynein. These observations imply that posttranslational nitrotyrosination of α-tubulin invokes conformational changes, either directly or via allosteric interactions, in the surface-exposed carboxyl terminus of α-tubulin that compromises the function of this critical domain in regulating microtubule organization and binding of motor- and microtubule-associated proteins. Collectively, these observations illustrate a mechanism whereby free NO2Tyr can impact deleteriously on cell function under pathological conditions encompassing reactive nitrogen species production. The data also yield further insight into the role that the α-tubulin tyrosination/detyrosination cycle plays in microtubule function.

Original languageEnglish (US)
Pages (from-to)6365-6370
Number of pages6
JournalProceedings of the National Academy of Sciences of the United States of America
Volume96
Issue number11
DOIs
StatePublished - 1999
Externally publishedYes

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Tubulin
Microtubules
Nitric Oxide
Wounds and Injuries
Cytoplasmic Dyneins
Reactive Nitrogen Species
Proteins
Microtubule-Associated Proteins
Tyrosine
Catalytic Domain
Pathology
Amino Acids
Enzymes

ASJC Scopus subject areas

  • Genetics
  • General

Cite this

Microtubule dysfunction by posttranslational nitrotyrosination of α- tubulin : A nitric oxide-dependent mechanism of cellular injury. / Eiserich, J. P.; Estevez, A. G.; Bamberg, T. V.; Yao Zu Ye, Zu Ye; Chumley, P. H.; Beckman, J. S.; Freeman, B. A.

In: Proceedings of the National Academy of Sciences of the United States of America, Vol. 96, No. 11, 1999, p. 6365-6370.

Research output: Contribution to journalArticle

Eiserich, J. P. ; Estevez, A. G. ; Bamberg, T. V. ; Yao Zu Ye, Zu Ye ; Chumley, P. H. ; Beckman, J. S. ; Freeman, B. A. / Microtubule dysfunction by posttranslational nitrotyrosination of α- tubulin : A nitric oxide-dependent mechanism of cellular injury. In: Proceedings of the National Academy of Sciences of the United States of America. 1999 ; Vol. 96, No. 11. pp. 6365-6370.
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