New challenges in pseudopodial proteomics by a laser-assisted cell etching technique.
New challenges in pseudopodial proteomics by a laser-assisted cell etching technique.
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DOI：
10.1016/j.bbapap.2014.10.009
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发表时间：
2015-06
期刊：
影响因子：
--
通讯作者：
T. Mimae;A. Ito
中科院分区：
文献类型：
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作者：
T. Mimae;A. Ito
伪足是一种向更高浓度的趋化物质延伸的腹膜突起，在细胞迁移和癌细胞侵袭中发挥关键作用。癌症，包括癌症和肉瘤，当它们侵犯周围结构和其他器官时，就会危及生命。了解侵袭性的分子基础对于消除癌症很重要。因此，确定伪足成分将有助于深入了解肿瘤细胞侵袭性的潜在机制，并提供潜在的生物标志物和治疗靶点。利用蛋白质组学方法对伪足的组成进行了广泛的研究。各种形式，包括基于凝胶和基于质谱学的方法，已经被用于伪足蛋白质组学。我们的研究小组最近建立了一种使用准分子激光脉冲选择性采集伪足的新方法，并成功地鉴定了一些新的伪足成分。在这里，我们总结了传统的蛋白质组学程序，并描述了我们的新的准分子激光辅助方法，特别强调了用于分离伪足的方法的差异。此外，我们还讨论了准分子激光介导的细胞消融在蛋白质组学应用中的理论背景。利用准分子激光辅助的方法，我们发现肌动蛋白结合接头蛋白α-Parvin定位于伪足，并与乳腺癌的侵袭性有关。我们的结果清楚地表明，准分子激光辅助细胞刻蚀是一种有用的伪足蛋白质组学技术。本文是题为：医学蛋白质组学特刊的一部分。
Pseudopodia are ventral membrane protrusions that extend toward higher concentrations of chemoattractants and play key roles in cell migration and cancer cell invasion. Cancers, including carcinoma and sarcoma, become life threatening when they invade surrounding structures and other organs. Understanding the molecular basis of invasiveness is important for the elimination of cancers. Thus, determining the pseudopodial composition will offer insights into the mechanisms underlying tumor cell invasiveness and provide potential biomarkers and therapeutic targets. Pseudopodial composition has been extensively investigated by using proteomic approaches. A variety of modalities, including gel-based and mass spectrometry-based methods, have been employed for pseudopodial proteomics. Our research group recently established a novel method using excimer laser pulses to selectively harvest pseudopodia, and we successfully identified a number of new pseudopodial constituents. Here, we summarized the conventional proteomic procedures and describe our new excimer laser-assisted method, with a special emphasis on the differences in the methods used to isolate pseudopodia. In addition, we discussed the theoretical background for the use of excimer laser-mediated cell ablation in proteomic applications. Using the excimer laser-assisted method, we showed that alpha-parvin, an actin-binding adaptor protein, is localized to pseudopodia, and is involved in breast cancer invasiveness. Our results clearly indicate that excimer laser-assisted cell etching is a useful technique for pseudopodial proteomics. This article is part of a Special Issue entitled: Medical Proteomics.