Demonstration of microtubule-like structures formed with (-)-rhazinilam from purified tubulin outside of cells and a simple tubulin-based assay for evaluation of analog activity Journal Article
| Authors: | Edler, M. C.; Yang, G.; Katherine Jung, M.; Bai, R.; Bornmann, W. G.; Hamel, E. |
| Article Title: | Demonstration of microtubule-like structures formed with (-)-rhazinilam from purified tubulin outside of cells and a simple tubulin-based assay for evaluation of analog activity |
| Abstract: | (-)-Rhazinilam was spontaneously generated from a natural product during isolation. In cultured cells, it causes microtubule bundle formation, like those caused by paclitaxel. With tubulin, (-)-rhazinilam causes formation of an aberrant spiral polymer. Using glutamate and GTP, we developed an assay for spiral formation and applied it to 17 new (±)-rhazinilam analogs with either a modified side chain or a different size D ring. There was reasonable correlation between spiral formation and inhibition of human MCF-7 breast carcinoma cell growth. Only one side chain analog was as active as (±)-rhazinilam. During these studies, we observed that omitting GTP from the reaction mixture caused a major change in the morphology of the (-)-rhazinilam-induced polymer, with half the observed polymer being microtubule-like and half being spirals. This mixed polymer slowly disassembled at 0 °C, but there was no apparent difference in the lability of the microtubules versus the spirals. |
| Keywords: | unclassified drug; human cell; antineoplastic agent; cell proliferation; animals; protein binding; drug structure; cell line, tumor; drug synthesis; structure activity relation; vinblastine; cancer inhibition; cell culture; protein purification; (-)-rhazinilam; glutamate; gtp; mcf-7 human breast carcinoma cells; microtubule morphology; rhazinilam analogs; sedimentation assay; spiral morphology of tubulin polymer; tubulin; glutamic acid; guanosine triphosphate; rhazinilam; cell strain mcf 7; drug determination; microtubule; temperature measurement; alkaloids; antimitotic agents; centrifugation; glutamates; indolizines; lactams; microtubules |
| Journal Title: | Archives of Biochemistry and Biophysics |
| Volume: | 487 |
| Issue: | 2 |
| ISSN: | 0003-9861 |
| Publisher: | Elsevier Inc. |
| Date Published: | 2009-07-15 |
| Start Page: | 98 |
| End Page: | 104 |
| Language: | English |
| DOI: | 10.1016/j.abb.2009.05.014 |
| PUBMED: | 19497297 |
| PROVIDER: | scopus |
| PMCID: | PMC2726660 |
| DOI/URL: | |
| Notes: | --- - "Cited By (since 1996): 1" - "Export Date: 30 November 2010" - "CODEN: ABBIA" - "Source: Scopus" |
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