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Industry article

Anti-CRISPR protein mediated degradation of Cas9 in human cells

Authors : Tacca, Luisa Arake de (2023)

Bacteriophages encode anti-CRISPR (Acr) proteins that inactivate CRISPR-Cas bacterial8 immune systems, allowing successful invasion, replication, and prophage integration. Acr9 proteins inhibit CRISPR-Cas systems using a wide variety of mechanisms. AcrIIA1 is encoded by10 numerous phages and plasmids, binds specifically to the Cas9 HNH domain, and was the first11 Acr discovered to inhibit SpyCas9. Here we report the observation of AcrIIA1-induced degradation12 ...

Journal article

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

Authors : Makarov, Dmytro (2023)

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 neuroblas...

Journal article

Novel human cell expression method reveals the role and prevalence of posttranslational modification in non-muscle tropomyosins

Authors : Carman, Peter J. (2021)

Biochemical studies require large protein quantities, which are typically obtained using bacterial expression. However, the folding machinery of bacteria is inadequate for many mammalian proteins, which additionally undergo posttranslational modifications (PTMs) that bacteria, yeast, or insect cells cannot perform. Many proteins also require native N- and C-termini and cannot tolerate extra tag amino acids for function. Tropomyosin, a coiled coil ...

Journal Article

SNM1A (DCLRE1A) is crucial for efficient repair of complex DNA breaks in human cells

Authors : Swift, Lonnie P. (2023)

DNA double-strand breaks (DSBs) are one of the most cytotoxic forms of damage, with even small number of unrepaired breaks being potentially lethal1. In human cells, the majority of DSBs are resolved by one of two pathways, that is non-homologous end-joining (active from G1- through to the G2-phase of the cell cycle) or homologous recombination (which is activated as cells traverse S-phase)2. Prior to completion of DSB repair,...