MATRIX-ASSISTED LASER DESORPTION/IONIZATION MASS-SPECTROMETRY (MALDI-MS) OF MEMBRANE-PROTEINS AND NONCOVALENT COMPLEXES
MATRIX-ASSISTED LASER DESORPTION/IONIZATION MASS-SPECTROMETRY (MALDI-MS) OF MEMBRANE-PROTEINS AND NONCOVALENT COMPLEXES
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DOI：
10.1002/jms.1190301012
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发表时间：
1995-10-01
影响因子：
2.3
通讯作者：
GALLA, HJ
中科院分区：
文献类型：
--
作者：
ROSINKE, B;STRUPAT, K;GALLA, HJ
基质辅助激光解吸/电离（MALDI）质谱和方法，以提高其质量的报告为三个疏水性，膜结合蛋白质：孔蛋白从大肠杆菌，细菌视紫红质盐杆菌和胆固醇酯酶从荧光假单胞菌。已经测试了几种常用的UV和IR基质。此外，敏感性的MALDI质谱的各种中性和离子洗涤剂，已知通常降低质量的MALDI质谱，已被系统地测试，对于孔蛋白，由三个相同的非共价键结合的亚基，一个新的样品制备的报告，导致在完整的四级蛋白质结构的解吸。这导致更好地理解给定分析物嵌入主体基质晶体的方式。
Matrix-assisted laser desorption/ionization (MALDI) mass spectra and methods to improve their quality are reported for three hydrophobic, membrane-bound proteins: porin from Escherichia coli, bacteriorhodopsin from Halobacterium salinarium and cholesterolesterase from Pseudomonas fluorescens. Several commonly used UV and IR matrices have been tested. In addition, the susceptibility of MALDI mass spectrometry to various neutral and ionic detergents, known usually to degrade the quality of MALDI mass spectra, has been tested systematically, For porin, consisting of three identical non-covalently bound subunits, a new sample preparation is reported, resulting in the desorption of the intact quaternary protein structure. This leads to a better understanding of the way a given analyte is embedded into the host matrix crystals.