Kim Sung-Kyoung, Rose Rebecca, Choksawangkarn Waeowalee, Graham Lauren, Hu Junkai, Fenselau Catherine, Lee Sang Bok
Department of Chemistry and Biochemistry, University of Maryland, College Park, MD, 20742.
Department of Chemistry and Biochemistry, University of Maryland, College Park, MD, 20742 ; Graduate School of Nanoscience and Technology, Korea Advanced Institute of Science and Technology, Daejeon, 305-701, Republic of Korea.
J Nanopart Res. 2013 Dec 1;15(12):2133. doi: 10.1007/s11051-013-2133-0.
A study is reported on the effect of nanowire density on the ease of pellicle formation and the enrichment of plasma membrane proteins for analysis by mass spectrometry. An optimized synthesis is reported for iron silicate nanowires with a narrow size range of 900 ±400 nm in length and 200 nm diameter. The nanowires were coated with AlO and used to form pellicles around suspended multiple myeloma cells, which acted as a model for cells recovered from tissue samples. Lighter alumina-coated silica nanowires were also synthesized (Kim et al. 2013), which allowed a comparison of the construction of the two pellicles and of the effect of nanowire density on plasma membrane enrichment. Evidence is offered that the dense nanowire pellicle does not crush or distort these mammalian cells. Finally, the pellicles were incorporated into a mass-spectrometry-based proteomic workflow to analyze transmembrane proteins in the plasma membrane. In contrast to a prior comparison of the effect of density with nanoparticles pellicles (Choksawangkarn et al. 2013), nanowire density was not found to significantly affect the enrichment of the plasma membrane. However, nanowires with a favorable aspect for pellicle formation are more easily and reliably produced with iron silicate than with silica. Additionally, the method for pellicle formation was optimized through the use of iron silicate nanowires (ISNW), which is crucial to the improvement of PM protein enrichment and analysis.
本文报道了一项关于纳米线密度对薄膜形成难易程度以及对质膜蛋白富集以用于质谱分析的影响的研究。报道了一种优化的硅酸铁纳米线合成方法,其长度尺寸范围狭窄,为900±400纳米,直径为200纳米。这些纳米线涂覆有氧化铝,并用于在悬浮的多发性骨髓瘤细胞周围形成薄膜,这些细胞作为从组织样本中回收的细胞的模型。还合成了较轻的氧化铝包覆的二氧化硅纳米线(Kim等人,2013年),这使得能够比较两种薄膜的构建以及纳米线密度对质膜富集的影响。有证据表明,密集的纳米线薄膜不会挤压或扭曲这些哺乳动物细胞。最后,将这些薄膜纳入基于质谱的蛋白质组学工作流程中,以分析质膜中的跨膜蛋白。与之前对纳米颗粒薄膜密度影响的比较(Choksawangkarn等人,2013年)不同,未发现纳米线密度对质膜富集有显著影响。然而,与二氧化硅相比,硅酸铁更易于且可靠地生产出有利于薄膜形成的纳米线。此外,通过使用硅酸铁纳米线(ISNW)优化了薄膜形成方法,这对改善质膜蛋白富集和分析至关重要。