3‐Azido‐L‐tyrosine as a photoinhibitor of tubulin:tyrosine ligase

[摘要]

We have synthesized the photoactivatable probes 3-azido-L-tyrosine andp-azido-L-phenylalanine and studied their capacity to inhibit the incorporation of[³H]tyrosine into tubulin catalyzed by tubulin:tyrosine ligase. Without illumination, only 3-azido-L-tyrosine reversibly inhibits the enzyme. Upon illumination, both reagents irreversibly photoinactivate the enzyme in a similar way. The ligase can be protected against photoinactivation by reversibly blocking essential thiol groups with pCMB during illumination.

[学科分类] 生物化学/生物物理

[关键词] Tubulin;Tubulin:tyrosine ligase;Posttranslational modification;3-Azido-L-tyrosine;Photoinaetivation;Thiol group;TTLase;tubulin:tyrosine ligase;Tub;tubulin;NH2Tyr;3-amino-L-tyrosine;Na3Tyr;3-azido-L-tyrosine;NH2Phe;p-amino-L-phenylalanine;N3Phe;p-azido-L-phenylalanine;pCMB;p-chloromercuribenzoic acid;DTE;dithioerythritol;DTNB (Ellman's reagent);5;5'-dithiobis-(2-nitrobenzoic acid);Mes;2-(N-morpholino)ethanesulfonic acid;ME;β-mercaptoethanol