Broad substrate tolerance of tubulin tyrosine ligase enables one-step site-specific enzymatic protein labeling.

Broad substrate tolerance of tubulin tyrosine ligase enables one-step site-specific enzymatic protein labeling† †The technology described in this manuscript is part of a pending patent application. ‡ ‡Electronic supplementary information (ESI) available: Supporting figures, synthetic details, HPLC and MS spectra, ubiquitin labeling. See DOI: 10.1039/c7sc00574a Click here for additional data file.

Tubulin-tyrosine ligase has a binding site on beta-tubulin: a two- domain structure of the enzyme

Tubulin-tyrosine ligase and alpha beta-tubulin form a tight complex which is conveniently monitored by glycerol gradient centrifugation. Using two distinct ligase monoclonal antibodies, several subunit-specific tubulin monoclonal antibodies, and chemical cross-linking, a ligase-binding site was identified on beta-tubulin. This site is retained when the carboxy-terminal domains of both tubulin s...

Tubulin Tyrosylation in uiuo and Changes Accompanying Differentiation of Cultured Neuroblastoma-Glioma

Changes in a posttranslational modification of tubulin, which accompany differentiation, have been studied in neuroblastoma-glioma hybrid cultured cells. The modification consists of the reversible enzymatic addition of a tyrosine to the COOH terminus of the (Y chain. Cytoplasmic tubulin purified from undifferentiated cells resembled that from adult mammalian brain in that half was in a form wh...

Characterization of the tubulin-tyrosine ligase

The sequence of tubulin-tyrosine ligase (TTL), the enzyme catalyzing the ATP-dependent posttranslational addition of a tyrosine to the carboxyterminal end of detyrosinated alpha-tubulin, has been determined. TTL from bovine and porcine brain was purified by immunoaffinity chromatography and extensively characterized by protein sequencing. Oligonucleotides derived from the protein sequence were ...

Parthenolide inhibits tubulin carboxypeptidase activity.

Microtubules are centrally involved in cell division, being the principal components of mitotic spindle. Tubulin, the constituent of microtubules, can be cyclically modified on its alpha-subunit by enzymatic removal of the COOH-terminal tyrosine residue by an ill-defined tubulin carboxypeptidase (TCP) and its readdition by tubulin tyrosine ligase (TTL). We and others have previously shown that ...