Comparison of nanowire pellicles for plasma membrane enrichment: coating nanowires on cell
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Comparison of nanowire pellicles for plasma membrane enrichment: coating nanowires on cell Sung-Kyoung Kim • Rebecca Rose • Waeowalee Choksawangkarn • Lauren M. Graham • Junkai Hu • Catherine Fenselau • Sang Bok Lee
Received: 28 August 2013 / Accepted: 14 November 2013 / Published online: 30 November 2013 Ó Springer Science+Business Media Dordrecht 2013
Abstract This study is reported on the effect of nanowire density on the ease of pellicle formation and the enrichment of plasma membrane (PM) proteins for analysis by mass spectrometry. An optimized synthesis is reported for iron silicate nanowires (ISNW) with a narrow size range of 900 ± 400 nm in length and 200 nm diameter. The nanowires were coated with Al2O3 and used to form pellicles around suspended multiple myeloma cells, which acted as a model for cells recovered from tissue samples. Lighter aluminacoated silica nanowires were also synthesized (Kim et al., doi:10.2217/NNM.13.96, 2013), which allowed a comparison of the construction of the two pellicles and of the effect of nanowire density on PM enrichment. Evidence is offered that the dense nanowire pellicle does not crush or distort these mammalian
Sung-Kyoung Kim and Rebecca Rose have contributed equally to this study. S.-K. Kim R. Rose W. Choksawangkarn L. M. Graham J. Hu C. Fenselau (&) S. B. Lee (&) Department of Chemistry and Biochemistry, University of Maryland, College Park, MD 20742, USA e-mail: [email protected] S. B. Lee e-mail: [email protected] S. B. Lee Graduate School of Nanoscience and Technology, Korea Advanced Institute of Science and Technology, Taejon 305-701, Republic of Korea
cells. Finally, the pellicles were incorporated into a mass spectrometry-based proteomic workflow to analyze transmembrane proteins in the PM. In contrast to a prior comparison of the effect of density with nanoparticles pellicles (Choksawangkarn et al. J Proteome Res 45512:1134–1141, 2013), nanowire density was not found to significantly affect the enrichment of the PM. However, nanowires with a favorable aspect for pellicle formation are more easily and reliably produced with iron silicate than with silica. In addition, the method for pellicle formation was optimized through the use of ISNW, which is crucial to the improvement of PM protein enrichment and analysis. Keywords Plasma membrane enrichment Nanowire pellicles Nanowire cell coating Silica nanowires Iron silicate nanowires
Introduction Due to their important role in communication and interactions between cells and their environment, plasma membrane (PM) proteins rank high among protein targets for the development of novel drugs (Overington et al. 2006). The successful study of PM proteins is critical to conducting pharmaceutical research, developing disease diagnostics, and elucidating fundamental biochemistry (Wu and Yates 2003). The most commonly used methods take advantage of the hydrophobicity of PM proteins
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(Speers and Wu 2007), or their accessibility to external chemical or physical probe
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