Norris Jeremy L, Porter Ned A, Caprioli Richard M
Mass Spectrometry Research Center, Department of Chemistry, Vanderbilt University, 465 21st Avenue South, Medical Research Building 3, Room 9160, Nashville, Tennessee 37232-8575, USA.
Anal Chem. 2003 Dec 1;75(23):6642-7. doi: 10.1021/ac034802z.
Detergents have been used to enhance the solubility of hydrophobic biomolecules for decades. Despite the widespread use of detergents in biochemistry, the presence of these molecules often complicates further analysis by mass spectrometry. This study presents a solution to this problem by outlining a method utilizing a novel cleavable detergent, 3-[3-(1,1-bisalkyloxyethyl)pyridin-1-yl]propane-1-sulfonate (PPS). This detergent can be used to extract protein contained within the interior of the cell by disrupting cell membranes. Once the proteins are free from the cell, PPS also assists in protein solubilization by shielding the hydrophobic regions of the newly extracted protein from unfavorable interactions with water. The added advantage of PPS over conventional detergents such as sodium dodecyl sulfate or n-octylglucoside is that the detergent properties that interfere with MALDI mass spectrometry can be eliminated prior to analysis. PPS was found to improve sensitivity in MALDI analyses of both soluble proteins and membrane proteins without degrading spectral quality. The virtues of this strategy were applied to whole cell extracts. Analysis of these extracts resulted in an overall increase in both the number of peaks observed and overall signal intensity.
几十年来，洗涤剂一直被用于提高疏水性生物分子的溶解度。尽管洗涤剂在生物化学领域广泛使用，但这些分子的存在常常使质谱进一步分析变得复杂。本研究提出了一种解决该问题的方法，概述了一种利用新型可裂解洗涤剂3-[3-(1,1-双烷氧基乙基)吡啶-1-基]丙烷-1-磺酸盐（PPS）的方法。这种洗涤剂可通过破坏细胞膜来提取细胞内部包含的蛋白质。一旦蛋白质从细胞中释放出来，PPS还通过保护新提取蛋白质的疏水区域免受与水的不利相互作用来协助蛋白质溶解。与传统洗涤剂如十二烷基硫酸钠或正辛基葡萄糖苷相比，PPS的额外优势在于其干扰基质辅助激光解吸电离质谱分析的洗涤剂特性可在分析前消除。研究发现，PPS在不降低光谱质量的情况下提高了可溶性蛋白质和膜蛋白的基质辅助激光解吸电离分析的灵敏度。该策略的优点应用于全细胞提取物。对这些提取物的分析导致观察到的峰数量和总体信号强度均全面增加。
