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Here, we report that the N-terminal intrinsically disordered region (IDR) of Kp60 is an essential bifunctional module for severing activity. Using solid-state NMR (ssNMR), we identified a microtubule-binding domain (MTBD) within the IDR which directly engages the lattice to guide substrate anchoring. Surprisingly, the same IDR also enables Kp60 phase separation. Our ssNMR analyses of the condensed phase pinpointed three discrete interaction regions within the IDR which cooperatively orchestrate this multivalent assembly. Importantly, Kp60 phase separation was associated with markedly increased ATPase activity, supporting a functional link between condensation and enzymatic activation. Using function-separation-mutation analyses, our studies show both microtubule binding and condensation are essential for severing: microtubule targeting provides spatial specificity, while condensation provides the catalytic threshold through local enrichment. Thus, the Kp60 IDR constitutes a bifunctional interface that integrates substrate recognition with enzymatic robustness, thereby establishing phase separation as a fundamental and efficient regulator of microtubule dynamics and cytoskeletal remodeling.<\/jats:p>","DOI":"10.1021\/jacs.6c15214","type":"journal-article","created":{"date-parts":[[2026,10,23]],"date-time":"2026-10-23T16:02:13Z","timestamp":1792771333000},"source":"Crossref","is-referenced-by-count":0,"title":["Solid-State NMR Reveals\na Bifunctional Disordered\nLinker That Couples Katanin Microtubule Anchoring to Condensate-Activated\nSevering"],"prefix":"10.1021","author":[{"given":"Panpan","family":"Xu","sequence":"first","affiliation":[{"name":"University of Science and Technology of China , , ,","place":["Hefei, China, 230027"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Wenping","family":"Hu","sequence":"additional","affiliation":[{"name":"University of Science and Technology of China , , ,","place":["Hefei, China, 230027"]},{"name":"University of Science and Technology of China , , ,","place":["Hefei, China, 230027"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Juan","family":"Li","sequence":"additional","affiliation":[{"name":"University of Science and Technology of China , , ,","place":["Hefei, China, 230027"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-0024-1096","authenticated-orcid":true,"given":"Chaowei","family":"Shi","sequence":"additional","affiliation":[{"name":"University of Science and Technology of China , , ,","place":["Hefei, China, 230027"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Xuebiao","family":"Yao","sequence":"additional","affiliation":[{"name":"University of Science and Technology of China , , ,","place":["Hefei, China, 230027"]},{"name":"University of Science and Technology of China , , ,","place":["Hefei, China, 230027"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Xing","family":"Liu","sequence":"additional","affiliation":[{"name":"University of Science and Technology of China , , ,","place":["Hefei, China, 230027"]},{"name":"University of Science and Technology of China , , ,","place":["Hefei, China, 230027"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-6874-4925","authenticated-orcid":true,"given":"ShengQi","family":"Xiang","sequence":"additional","affiliation":[{"name":"University of Science and Technology of China , , ,","place":["Hefei, China, 230027"]}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"316","published-online":{"date-parts":[[2026,10,23]]},"reference":[{"issue":"5991","key":"2026102312020559200_cit1","doi-asserted-by":"publisher","first-page":"237","DOI":"10.1038\/312237a0","article-title":"Dynamic instability of microtubule growth","volume":"312","author":"Mitchison","year":"1984","journal-title":"Nature"},{"issue":"12","key":"2026102312020559200_cit2","doi-asserted-by":"publisher","first-page":"777","DOI":"10.1038\/s41580-021-00399-x","article-title":"Regulation of microtubule\ndynamics, mechanics and function\nthrough the growing tip","volume":"22","author":"Gudimchuk","year":"2021","journal-title":"Nat. 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