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Custom Vector Construction> | … For the generation of cell lines, the full-length katanin open reading frame (RefSeq NM_0070444) was purchased from GenScript and subcloned into the pcDNA5.0/FRT/TO vector (Thermo Fisher) with an N-terminal EGFP tag using KpnI and NotI restriction sites. Vectors for … | Get A Quote |
The structural conservation across the AAA (ATPases associated with diverse cellular activities) protein family makes designing selective chemical inhibitors challenging. Here, we identify a triazolopyridine-based fragment that binds the AAA domain of human katanin, a microtubule-severing protein. We have developed a model for compound binding and designed ASPIR-1 (allele-specific, proximity-induced reactivity-based inhibitor-1), a cell-permeable compound that selectively inhibits katanin with an engineered cysteine mutation. Only in cells expressing mutant katanin does ASPIR-1 treatment increase the accumulation of CAMSAP2 at microtubule minus ends, confirming specific on-target cellular activity. Importantly,... More