PT  - JOURNAL ARTICLE
AU  - Dmytro Makarov
AU  - Pavel Kielkowski
TI  - Chemical Proteomics Reveals Protein Tyrosination Extends Beyond the Alpha-Tubulins in Human Cells
AID  - 10.1101/2022.07.02.498566
DP  - 2022 Jan 01
TA  - bioRxiv
PG  - 2022.07.02.498566
4099  - http://biorxiv.org/content/early/2022/07/03/2022.07.02.498566.short
4100  - http://biorxiv.org/content/early/2022/07/03/2022.07.02.498566.full
AB  - Tubulin detyrosination-tyrosination cycle regulates the stability of microtubules. Thus far described on α-tubulins, the tyrosination level is maintained by a single tubulin-tyrosine ligase (TTL). However, the precise dynamics and tubulin isoforms which undergo (de)tyrosination in neurons are unknown. Here, we exploit the substrate promiscuity of the TTL to introduce an O-propargyl-L-tyrosine in neuroblastoma cells and neurons. Mass spectrometry-based chemical proteomics in neuroblastoma cells using the O-propargyl-L-tyrosine probe revealed previously discussed tyrosination of TUBA4A, MAPRE1, and other non-tubulin proteins. This finding was further corroborated in differentiating neurons. Together we present the method for tubulin tyrosination profiling in living cells. Our results show that detyrosination-tyrosination is not restricted to α-tubulins with coded C-terminal tyrosine and is thus involved in fine-tuning of the tubulin and non-tubulin proteins during neuronal differentiation.Competing Interest StatementThe authors have declared no competing interest.