Publications

• Tubulin tyrosine ligase structure reveals adaptation of an ancient fold to bind and modify tubulin.

  Szyk A., Deaconescu A.M., Piszczek G., Roll-Mecak A.

  Tubulin tyrosine ligase (TTL) catalyzes the post-translational C-terminal tyrosination of α-tubulin. Tyrosination regulates recruitment of microtubule-interacting proteins. TTL is essential. Its loss causes morphogenic abnormalities and is associated with cancers of poor prognosis. We present the ... >> More

  Tubulin tyrosine ligase (TTL) catalyzes the post-translational C-terminal tyrosination of α-tubulin. Tyrosination regulates recruitment of microtubule-interacting proteins. TTL is essential. Its loss causes morphogenic abnormalities and is associated with cancers of poor prognosis. We present the first crystal structure of TTL (from Xenopus tropicalis), defining the structural scaffold upon which the diverse TTL-like family of tubulin-modifying enzymes is built. TTL recognizes tubulin using a bipartite strategy. It engages the tubulin tail through low-affinity, high-specificity interactions, and co-opts what is otherwise a homo-oligomerization interface in structurally related ATP grasp-fold enzymes to form a tight hetero-oligomeric complex with the tubulin body. Small-angle X-ray scattering and functional analyses reveal that TTL forms an elongated complex with the tubulin dimer and prevents its incorporation into microtubules by capping the tubulin longitudinal interface, possibly modulating the partition of tubulin between monomeric and polymeric forms. << Less

  Nat Struct Mol Biol 18:1250-1258(2011) [PubMed] [EuropePMC]

• The carboxy-terminal peptide of detyrosinated alpha tubulin provides a minimal system to study the substrate specificity of tubulin-tyrosine ligase.

  Rudiger M., Wehland J., Weber K.

  The ATP-dependent tubulin-tyrosine ligase (TTL) restores the carboxy-terminal tyrosine of alpha tubulin in alpha beta tubulin that has been previously detyrosinated. Here we show that the carboxy-terminal tetradecapeptide of detyrosinated alpha tubulin is used by TTL as a substrate, albeit at 50-f ... >> More

  The ATP-dependent tubulin-tyrosine ligase (TTL) restores the carboxy-terminal tyrosine of alpha tubulin in alpha beta tubulin that has been previously detyrosinated. Here we show that the carboxy-terminal tetradecapeptide of detyrosinated alpha tubulin is used by TTL as a substrate, albeit at 50-fold lower efficiency than alpha beta tubulin. The minimal system provided by the TTL/peptide combination mirrors the TTL/tubulin system in all aspects tested, and shows a pronounced substrate inhibition. Synthetic peptides varying in length and/or containing single amino acid replacements were used to analyze the TTL specificity for the carboxy-terminal sequence of detyrosinated alpha tubulin. Peptides ending like alpha tubulin with the sequence Gly-Glu-Glu are optimally tyrosinated once a peptide length of 12 residues is reached. Position -1 of this recognition sequence, to which the tyrosine is added, must be glutamic acid. Position -2 accepts only an acidic amino acid but glutamic acid is by far preferred over aspartic acid. These results explain why a subpopulation of brain alpha tubulin, which ends with the sequence Gly-Glu, is not tyrosinated by TTL. The carboxy-terminal dodecapeptide of brain alpha tubulin with its polyglutamyl side-chain on position -6 shows the same substrate activity as the corresponding synthetic peptide lacking the side-chain. We discuss the substrate specificity of TTL for different alpha tubulins and speculate why tubulin is a better substrate than the optimal peptide covering the carboxy-terminal of detyrosinated alpha tubulin. << Less

  Eur J Biochem 220:309-320(1994) [PubMed] [EuropePMC]

• Structural basis of tubulin tyrosination by tubulin tyrosine ligase.

  Prota A.E., Magiera M.M., Kuijpers M., Bargsten K., Frey D., Wieser M., Jaussi R., Hoogenraad C.C., Kammerer R.A., Janke C., Steinmetz M.O.

  Tubulin tyrosine ligase (TTL) catalyzes the post-translational retyrosination of detyrosinated α-tubulin. Despite the indispensable role of TTL in cell and organism development, its molecular mechanism of action is poorly understood. By solving crystal structures of TTL in complex with tubulin, we ... >> More

  Tubulin tyrosine ligase (TTL) catalyzes the post-translational retyrosination of detyrosinated α-tubulin. Despite the indispensable role of TTL in cell and organism development, its molecular mechanism of action is poorly understood. By solving crystal structures of TTL in complex with tubulin, we here demonstrate that TTL binds to the α and β subunits of tubulin and recognizes the curved conformation of the dimer. Biochemical and cellular assays revealed that specific tubulin dimer recognition controls the activity of the enzyme, and as a consequence, neuronal development. The TTL-tubulin structure further illustrates how the enzyme binds the functionally crucial C-terminal tail sequence of α-tubulin and how this interaction catalyzes the tyrosination reaction. It also reveals how TTL discriminates between α- and β-tubulin, and between different post-translationally modified forms of α-tubulin. Together, our data suggest that TTL has specifically evolved to recognize and modify tubulin, thus highlighting a fundamental role of the evolutionary conserved tubulin tyrosination cycle in regulating the microtubule cytoskeleton. << Less

  J Cell Biol 200:259-270(2013) [PubMed] [EuropePMC]

• Isolation and purification of tubulin tyrosine ligase.

  Wehland J., Schroder H.C., Weber K.

  Methods Enzymol 134:170-179(1986) [PubMed] [EuropePMC]

• Tubulin tyrosine ligase: protein and mRNA expression in developing rat skeletal muscle.

  Arregui C.O., Mas C.R., Argarana C.E., Barra H.S.

  Alpha tubulin can be post-translationally tyrosinated at the carboxy-terminus by a specific enzyme: tubulin tyrosine ligase. The expression of tubulin tyrosine ligase mRNA and protein during the development of rat skeletal muscle was examined in the present study. A portion of the coding region of ... >> More

  Alpha tubulin can be post-translationally tyrosinated at the carboxy-terminus by a specific enzyme: tubulin tyrosine ligase. The expression of tubulin tyrosine ligase mRNA and protein during the development of rat skeletal muscle was examined in the present study. A portion of the coding region of the rat ligase cDNA was isolated and sequenced. The nucleotide and amino acid sequences showed about 90% homology with previously reported porcine and bovine ligase sequences. In newborn rats, ligase mRNA and protein were highly expressed in skeletal muscle. During early postnatal development, however, both ligase mRNA and protein dropped down dramatically. Quantitative measurements revealed that ligase protein at postnatal day 20 represented only 10% or less of the level at postnatal day 1. Ligase mRNA expression was also examined during the myogenesis in vitro. A strong ligase mRNA signal was detected in both undifferentiated myoblasts and cross-striated, contractile myotubes. The present results suggest that, during muscle differentiation, ligase function may be regulated by the amount of available mRNA. The discrepancy in the ligase expression between the in vivo and in vitro myogenesis suggests that factors controlling the levels of mRNA in vivo are lost in vitro. << Less

  Dev. Growth Differ. 39:167-178(1997) [PubMed] [EuropePMC]