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Under sub-critical concentration conditions ([Tu-GTP]&amp;lt;5.3 \u03bcM), significant microtubule growth of limited length was observed on a proportion of the EGS-seeds by immuno-electron microscopy. A sensitive fluorescence assay for microtubule GDP production was developed for parallel assessment of GTP utilisation. This revealed a correlation between the detected microtubule growth and the production of tubulin-GDP, deriving from the shortening phase of the dynamic microtubules. This correlation was confirmed by the action of nocodazole, a specific inhibitor of microtubule assembly, that was found to abolish the GDP release. The variation of the GDP release with tubulin concentration (Jh(c) plot) was determined below the critical concentration (CC). The GDP production observed was consistent with the elongation of the observed seeded microtubules with an apparent rate constant of 1.5\u00d7106M\u22121 second\u22121 above a threshold of approximately 1 \u03bcM tubulin. The form of this Jh(c) plot for elongation below CC is reproduced by the Lateral Cap model for microtubule dynamic instability adapted for seeded assembly. The behaviour of the system is contrasted with that previously studied in the absence of detectable microtubule elongation (Caplow and Shanks (1990)J. Biol. Chem. 265, 8935-8941). The approach provides a means of monitoring microtubule dynamics at concentrations inaccessible to optical microscopy, and shows that essentially the same dynamic mechanisms apply at all concentrations. Numerical simulation of the subcritical concentration regime shows dynamic growth features applicable to the initiation of microtubule growth in vivo.<\/jats:p>","DOI":"10.1242\/jcs.109.11.2755","type":"journal-article","created":{"date-parts":[[2021,4,25]],"date-time":"2021-04-25T16:28:29Z","timestamp":1619368109000},"page":"2755-2766","source":"Crossref","is-referenced-by-count":10,"title":["Dynamic properties of nucleated microtubules: GTP utilisation in the sub-critical concentration regime"],"prefix":"10.1242","volume":"109","author":[{"given":"Martyn F.","family":"Symmons","sequence":"first","affiliation":[{"name":"Division of Physical Biochemistry, National Institute for Medical Research, Mill Hill, London NW7 1AA, UK"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Stephen R.","family":"Martin","sequence":"additional","affiliation":[{"name":"Division of Physical Biochemistry, National Institute for Medical Research, Mill Hill, London NW7 1AA, UK"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Peter M.","family":"Bayley","sequence":"additional","affiliation":[{"name":"Division of Physical Biochemistry, National Institute for Medical Research, Mill Hill, London NW7 1AA, UK"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"237","published-online":{"date-parts":[[1996,11,1]]},"reference":[{"key":"2024010323574476000_JOCES_109_11_2755C1","doi-asserted-by":"publisher","first-page":"589","DOI":"10.1016\/0092-8674(94)90496-0","article-title":"Deconstructing the microtubule organizing center","volume":"76","author":"Archer","year":"1994","journal-title":"Cell"},{"key":"2024010323574476000_JOCES_109_11_2755C2","doi-asserted-by":"publisher","first-page":"495","DOI":"10.1083\/jcb.111.2.495","article-title":"Individual microtubules in the axon consist of domains that differ in both composition and stability","volume":"111","author":"Baas","year":"1990","journal-title":"J. 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