School of Molecular Bioscience, University of Sydney, Sydney, NSW 2006, Australia.
Anal Chem. 2012 Apr 17;84(8):3725-30. doi: 10.1021/ac300291c. Epub 2012 Mar 28.
A matrix-assisted laser desorption ionization (MALDI) mass spectrometry-based approach is applied to identify active site domains within influenza neuraminidase that bind the antiviral inhibitors zanamivir (ZANA) and 2-deoxy-2,3-didehydro-N-acetylneuraminic acid (DANA). Combined data from the tryptic and Glu-C endoproteinase digests of neuraminidase-inhibitor complexes have identified binding peptides that contain the active site residues Arg118, Glu119, Arg156, Glu276, and Tyr406. The binding of these residues was confirmed from the analysis of available X-ray crystal structures. The ability to identify peptides within the active sites of proteins and likely binding residues provides both a rapid and relatively high throughput approach with which to screen protein-drug interactions by MALDI mass spectrometry.
基质辅助激光解吸电离（MALDI）质谱法被应用于鉴定流感神经氨酸酶中的活性位点结构域，这些结构域能够与抗病毒抑制剂扎那米韦（ZANA）和 2-脱氧-2,3-二脱氢-N-乙酰神经氨酸（DANA）结合。来自神经氨酸酶-抑制剂复合物的胰蛋白酶和 Glu-C 内切蛋白酶消化的综合数据，鉴定出了包含活性位点残基 Arg118、Glu119、Arg156、Glu276 和 Tyr406 的结合肽。这些残基的结合通过对现有 X 射线晶体结构的分析得到了证实。通过 MALDI 质谱法，能够在蛋白质的活性位点和可能的结合残基内鉴定出肽，这为筛选蛋白质-药物相互作用提供了一种快速且相对高通量的方法。
