Microtubule regulation by small molecules.

小分子的微管调节。

• 批准号: 7734779
• 负责人: Dan L Sackett
• 金额: $ 24.8万
• 依托单位: EUNICE KENNEDY SHRIVER NATIONAL INSTITUTE OF CHILD HEALTH & HUMAN DEVELOPMENT
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/7734779
• 关键词: Affect; Antimitotic Agents; Binding; Biological Assay; Cells; Cellular Morphology; Characteristics; Combretastatin; Cryoelectron Microscopy; Crystallization; Cytoplasm; Cytoskeleton; Disease; Endopeptidases; Fluorescence Microscopy; Generations; Human; Knowledge; Leishmania; Malignant Glioma; Maps; Marines; Microtubule Polymerization; Microtubules; Mitotic; Movement; Nitrosourea Compounds; Numbers; Parasites; Peptide Hydrolases; Peptides; Pharmacotherapy; Polymers; Post-Translational Protein Processing; Property; Proteasome Inhibitor; Protein Subunits; Proteins; Range; Regulation; Resolution; Role; Shapes; Source; Spectrum Analysis; Structure; Thalidomide; Tubulin; Yeasts; analog; analytical ultracentrifugation; chemotherapy; human disease; macrophage; marine natural product; migration; novel; physical property; prevent; small molecule; stathmin; trafficking

We have focused on antimitotic peptides, often of marine origin, because these are among the most potent anti-MT agents known, they have been synthesized and analogs are available, and because they induce the MT subunits to assume unusual and characteristic ring shapes. We are studying the structural and dynamic properties of these ring polymers by analytical ultracentrifugation, cryoelectron microscopy, fluorescence correlation spectroscopy, and protease mapping. The high stability and uniformity of these rings that our studies revealed have led us to attempt crystallization of these polymers to achieve atomic resolution of their structure. We are also examining the effects on microtubule polymerization of synthetic analogs of thalidomide and combretastatin A. We have also demonstrated the role of posttranslational modifications by deacetylases, and the effects of proteasome inhibitors on microtubule stability. We have identified a number of new modified peptides derived from the natural peptide tubulysin. Thes new peptides show a range of antimicrotubule activity in assays with purified proteins as well as in cells. In addition we have examined a new "second generation" of microtubule stabilizers, focusing on the natural compound peloruside. We have also shown that an old chemotherapy agent, a nitrosourea, may affect microtubule stability indirectly by altering the activity of the protein stathmin. This results in reduced migration and invasion by malignant glioma cells. We are seeking to identify small molecules that do not bind well with mammalian tubulin but do bind to parasite tubulin. The tubulin molecule is quite conserved evolutionarily, but differences do exist, and several molecules are known that can target, for example, yeast rather than mammalian tubulin or vice-versa. We are looking for molecules that will target Leishmania, the infectious cause of an important group of human diseases. We have identified several small molecules that show promise as selective agents, binding to Leishmania tubulin preferentially over mammalian tubulin, and preventing parasite multiplication inside human macrophage cells.

我们一直专注于抗有丝分裂多肽，通常来自海洋，因为它们是已知的最有效的抗MT药物之一，它们已经被合成和类似物可用，而且因为它们诱导MT亚单位呈现不寻常的和特征的环状。我们正在通过分析超速离心法、低温电子显微镜、荧光相关光谱和蛋白酶图谱来研究这些环聚合物的结构和动力学性质。我们的研究揭示了这些环的高度稳定性和一致性，这使得我们试图对这些聚合物进行结晶，以实现其结构的原子分辨。我们还在研究合成的沙利度胺和comretasteatin A的类似物对微管聚合的影响。我们还证明了脱乙酰酶的翻译后修饰的作用，以及蛋白酶体抑制剂对微管稳定性的影响。 我们已经从天然多肽Tubulysin中鉴定出一些新的修饰多肽。这些新的多肽在与纯化蛋白的检测中以及在细胞中都显示出一系列的抗微管活性。此外，我们还研究了新的“第二代”微管稳定剂，重点是天然化合物Peloruside。我们还表明，一种古老的化疗药物亚硝脲可能通过改变Stathmin蛋白的活性来间接影响微管的稳定性。这会减少恶性胶质瘤细胞的迁移和侵袭。 我们正在寻找不能很好地与哺乳动物微管蛋白结合但确实能与寄生虫微管蛋白结合的小分子。微管蛋白分子在进化上是相当保守的，但确实存在差异，并且已知有几个分子可以靶向例如酵母而不是哺乳动物的微管蛋白，反之亦然。我们正在寻找针对利什曼原虫的分子，利什曼原虫是人类一组重要疾病的感染原因。我们已经确定了几个小分子作为选择性试剂，优先与利什曼原虫微管蛋白结合，而不是哺乳动物微管蛋白，并防止寄生虫在人巨噬细胞内繁殖。

项目成果

Selective antimicrotubule activity of N1-phenyl-3,5-dinitro-N4,N4-di-n-propylsulfanilamide (GB-II-5) against kinetoplastid parasites.

N1-苯基-3,5-二硝基-N4,N4-二正丙基磺酰胺 (GB-II-5) 对动质体寄生虫的选择性抗微管活性。

• DOI: 10.1124/mol.64.6.1325
• 发表时间: 2003
• 期刊: Molecular pharmacology
• 影响因子: 3.6
• 作者: Werbovetz,KarlA;Sackett,DanL;Delfin,Dawn;Bhattacharya,Gautam;Salem,Manar;Obrzut,Tomasz;Rattendi,Donna;Bacchi,Cyrus
• 通讯作者: Bacchi,Cyrus

Intracellular proadrenomedullin-derived peptides decorate the microtubules and contribute to cytoskeleton function.

细胞内肾上腺髓质素原衍生肽装饰微管并有助于细胞骨架功能。

• DOI: 10.1210/en.2007-1763
• 发表时间: 2008
• 期刊: Endocrinology
• 影响因子: 4.8
• 作者: Sackett,DanL;Ozbun,Laurent;Zudaire,Enrique;Wessner,Lisa;Chirgwin,JohnM;Cuttitta,Frank;Martinez,Alfredo
• 通讯作者: Martinez,Alfredo