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Top‐down characterization of proteins and drug‐protein complexes using nanoelectrospray tandem mass spectrometry
Author(s) -
Mandal Rupasri,
Li XingFang
Publication year - 2005
Publication title -
rapid communications in mass spectrometry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.528
H-Index - 136
eISSN - 1097-0231
pISSN - 0951-4198
DOI - 10.1002/rcm.2268
Subject(s) - chemistry , tandem mass spectrometry , mass spectrometry , myoglobin , top down proteomics , peptide , chromatography , fragmentation (computing) , protein mass spectrometry , peptide mass fingerprinting , biochemistry , proteomics , operating system , computer science , gene
We report a ‘top‐down’ approach for characterization of proteins, and identification of binding sites in protein‐drug complexes using nanoelectrospray ionization hybrid quadrupole time‐of‐flight tandem mass spectrometry (nanoESI‐MS/MS). The efficiency of direct fragmentation of intact protein ions and the feasibility of this method were initially demonstrated using several well‐characterized proteins with different molecular weights including metallothionein (6126 Da), cytochrome c (horse, 12360 Da), myoglobin (horse, 16592 Da), and hemoglobin (human, 64453 Da). Simply varying collision energy without enzyme digestion and gel or LC separation generated a range of peptide fragments of these proteins. Over 80% of these peptide ions matched those in the SWISS‐PROT database with mass accuracy of 8 to 32 ppm with external calibration. This technique was further applied to fragment a cisplatin‐metallothionein complex to identify the binding sites, demonstrating a potential application in the study of drug‐protein binding. Copyright © 2005 John Wiley & Sons, Ltd.