O'Connell Paul A, Pinto Devanand M, Chisholm Ken A, MacRae Thomas H
Department of Biology, Dalhousie University, Halifax, Nova Scotia, Canada B3H 4J1.
Biochim Biophys Acta. 2006 May;1764(5):920-8. doi: 10.1016/j.bbapap.2006.03.003. Epub 2006 Apr 5.
The microtubule proteome encompasses tubulin and a diverse group of proteins which associate with tubulin upon microtubule formation. These proteins either determine microtubule organization and function or their activity is influenced by microtubule association. To characterize the microtubule proteome in Artemia franciscana, tubulin assembly was induced with taxol in vitro after 0 and 12 h of post-diapause development. Proteins obtained by extraction of microtubules with 0.5 M NaCl were electrophoresed in two-dimensional gels and analyzed by mass spectrometry. Fifty-five proteins were identified with 10 of these occurring at both developmental stages, and multiple isoforms were observed for some proteins of the Artemia proteome. Their functions include roles in membrane transport, metabolism, chaperoning and protein synthesis, thus reflecting physiological properties of encysted Artemia such as stress resistance and the ability to rapidly initiate post-diapause development. For example, chaperones may protect tubulin during encystment and facilitate folding in metabolically active embryos. Additionally, the interaction of metabolic enzymes with microtubules funnels reaction intermediates, potentially enhancing efficiency within biochemical processes. This study represents the first systematic characterization of a crustacean microtubule proteome. Although it is difficult to be certain that all protein associations documented herein occur in vivo, the results suggest how protein-protein interactions contribute to cytoplasmic organization while implying how Artemia embryos resist stress and remain capable of development once diapause terminates.
微管蛋白质组包含微管蛋白以及在微管形成时与微管蛋白相关联的多种蛋白质。这些蛋白质要么决定微管的组织和功能，要么其活性受微管关联的影响。为了表征卤虫的微管蛋白质组，在滞育后发育0小时和12小时后，用紫杉醇在体外诱导微管蛋白组装。用0.5M NaCl提取微管得到的蛋白质在二维凝胶上进行电泳，并通过质谱分析。鉴定出了55种蛋白质，其中10种在两个发育阶段均出现，并且在卤虫蛋白质组的一些蛋白质中观察到了多种同工型。它们的功能包括在膜运输、代谢、伴侣作用和蛋白质合成中的作用，从而反映了包囊卤虫的生理特性，如抗逆性和迅速启动滞育后发育的能力。例如，伴侣蛋白可能在包囊形成过程中保护微管蛋白，并促进代谢活跃胚胎中的折叠。此外，代谢酶与微管的相互作用汇集反应中间体，可能提高生化过程中的效率。这项研究代表了对甲壳类微管蛋白质组的首次系统表征。尽管很难确定本文记录的所有蛋白质关联都在体内发生，但结果表明了蛋白质-蛋白质相互作用如何促进细胞质组织，同时暗示了卤虫胚胎如何抵抗压力并在滞育结束后仍能发育。
