Wignall Sarah M, Gray Nathanael S, Chang Young-Tae, Juarez Lolita, Jacob Richard, Burlingame Al, Schultz Peter G, Heald Rebecca
Department of Molecular and Cell Biology, University of California, Berkeley, 311 Life Sciences Addition, Berkeley, California 94720, USA.
Chem Biol. 2004 Jan;11(1):135-46.
To identify novel proteins regulating the microtubule cytoskeleton, we screened a library of purine derivatives using mitotic spindle assembly in Xenopus egg extracts as an assay. Out of a collection of 1561 compounds, we identified 15 that destabilized microtubules without targeting tubulin directly, resulting in small spindles. Affinity chromatography with one compound, named diminutol, revealed a potential target as NQO1, an NADP-dependent oxidoreductase. A role for NQO1 in influencing microtubule polymerization was confirmed through inhibition studies using known inhibitors and immunodepletion. Therefore, this chemical approach has identified a novel factor required for microtubule morphogenesis and cell division.
为了鉴定调控微管细胞骨架的新型蛋白质，我们利用非洲爪蟾卵提取物中的有丝分裂纺锤体组装作为检测方法，对嘌呤衍生物文库进行了筛选。在1561种化合物中，我们鉴定出15种能使微管不稳定且不直接作用于微管蛋白的化合物，这些化合物导致纺锤体变小。用一种名为小纺锤体醇（diminutol）的化合物进行亲和层析，揭示了一个潜在靶点为NQO1，一种依赖NADP的氧化还原酶。通过使用已知抑制剂的抑制研究和免疫耗竭，证实了NQO1在影响微管聚合中的作用。因此，这种化学方法鉴定出了微管形态发生和细胞分裂所需的一种新因子。
