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Chemical Proteomics Reveals Protein Tyrosination Extends Beyond the Alpha-Tubulins in Human Cells

Dmytro Makarov, View ORCID ProfilePavel Kielkowski
doi: https://doi.org/10.1101/2022.07.02.498566
Dmytro Makarov
1LMU München, Department of Chemistry, Institute for Chemical Epigenetics – Munich (ICEM), Würmtalstrasse 201, 81375, Munich, Germany
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Pavel Kielkowski
1LMU München, Department of Chemistry, Institute for Chemical Epigenetics – Munich (ICEM), Würmtalstrasse 201, 81375, Munich, Germany
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  • ORCID record for Pavel Kielkowski
  • For correspondence: pavel.kielkowski@cup.lmu.de
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Abstract

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 Statement

The authors have declared no competing interest.

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The copyright holder for this preprint is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made available under a CC-BY-NC-ND 4.0 International license.
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Posted July 03, 2022.
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Chemical Proteomics Reveals Protein Tyrosination Extends Beyond the Alpha-Tubulins in Human Cells
Dmytro Makarov, Pavel Kielkowski
bioRxiv 2022.07.02.498566; doi: https://doi.org/10.1101/2022.07.02.498566
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Chemical Proteomics Reveals Protein Tyrosination Extends Beyond the Alpha-Tubulins in Human Cells
Dmytro Makarov, Pavel Kielkowski
bioRxiv 2022.07.02.498566; doi: https://doi.org/10.1101/2022.07.02.498566

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