کد مقاله کد نشریه سال انتشار مقاله انگلیسی نسخه تمام متن
7633617 1494706 2018 29 صفحه PDF دانلود رایگان
عنوان انگلیسی مقاله ISI
Ultra-high mass multimer analysis of protein-1a capping domains by a silicon nanomembrane detector
موضوعات مرتبط
مهندسی و علوم پایه شیمی شیمی آنالیزی یا شیمی تجزیه
پیش نمایش صفحه اول مقاله
Ultra-high mass multimer analysis of protein-1a capping domains by a silicon nanomembrane detector
چکیده انگلیسی
Conventional time of flight ion detectors are based on secondary electron multipliers encountering a significant loss in detection efficiency, sensitivity and resolution with protein mass above 50 kDa. In this work we employ a silicon nanomembrane detector in a Matrix-Assisted Laser Desorption/Ionization coupled to time of flight (MALDI-TOF) mass spectrometer. The operating principle relies on phonon-assisted field emission with excellent performance in the high mass range from 0.001-2 MDa. In addition to the analysis of standard proteins the nanomembrane detector (NMD) has the potential for the detection and structural investigation of complex macromolecular assemblies through non-covalent interactions. In order to investigate this hypothesis, the N-terminal capping/methyltransferase domain (CAP) of the Brome Mosaic Virus (BMV) 1a replication protein by MALDI-TOF-NMD is analyzed. The signals detected at the high m/z-ratios of 912.6/982.7 (× 103) and 1333.3 (× 103) could be modified species of CAP-tricta/tetractamer and the octadecamer. For the first time, the NMD is applied to detect biologically complex macromolecular protein assemblies. Hence, this technology overcomes the limitations of conventional TOF-detectors and increases the analytical range of MALDI-TOF. This technology will be a future alternative for the structural analysis of intact virus capsids that will complement other MS-based techniques such as native mass spectrometry.
ناشر
Database: Elsevier - ScienceDirect (ساینس دایرکت)
Journal: Journal of Proteomics - Volume 175, 20 March 2018, Pages 5-11
نویسندگان
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