Application of mass spectrometry for target identification and characterization

Abstract

Mass spectrometry‐based methodologies span the vast expanse of drug discovery. Both electrospray ionization (ESI) and matrix‐assisted laser desorption/ionization (MALDI) support proteomics‐based research projects by identifying proteins separated and isolated by polyacrylamide gel electrophoresis. MALDI‐MS‐based surface scanning of one‐dimensional isoelectric focusing gels, “virtual 2‐D gel electrophoresis,” represents a potentially high throughput means to map proteins and to determine protein profiles. Mass spectrometry can also be used to directly study the covalent and noncovalent interactions of drug molecules and biomolecule targets. Drug binding examples discussed include the binding of covalent and noncovalent inhibitors to src SH2 domain protein, and the interaction of aminoglycoside antibiotic neomycin and HIV Tat peptide‐TAR RNA. © 1999 John Wiley & Sons, Inc. Med Res Rev, 19, No. 4, 307–319, 1999.

Mass spectrometry captures off-target drug binding and provides mechanistic insights into the human metalloprotease ZMPSTE24

“…In conclusion, the mass spectrometry approaches that we applied in this study extend earlier studies of metal binding and drugs to soluble protein targets 40–44 and provide direct evidence for the binding of HIV protease inhibitors to the membrane protein ZMPSTE24, with and without zinc binding, and in the presence of detergent and lipid adducts. This is not only the first direct evidence of drug binding to ZMPSTE24 but our results are corroborated by the resistance to unfolding conferred by these binding events.…”

Targeting the HIV Trans‐Activation Responsive Region—Approaches Towards RNA‐Binding Drugs

“…Mass spectroscopy is another possible way to study the stoichiometry of aminoglycoside–TAR interactions 31. Published data indicate that one TAR molecule may be complexed by up to three neomycin molecules.…”

Mass spectrometry in high throughput analysis

“…In the field of drug development, high throughput chemistry represents a convergence of chemistry with biology, made possible by fundamental advances in automation, such as that of mass spectrometry [7][8][9][10][11][12][13][14][15][16][17][18][19]. High throughput technology has even been extended to proteomics, where mass spectrometry is being used as a tool in rapid protein identification [20][21][22].…”

“…The automation of MALDI and DIOS analyses is becoming increasingly important in proteomics and combinatorial chemistry [5,11,[20][21][22]. Typically, the instrument is designed to search for a signal from each sample well.…”

“…In the field of drug development, high throughput chemistry represents a convergence of chemistry with biology, made possible by fundamental advances in automation, such as that of mass spectrometry [7][8][9][10][11][12][13][14][15][16][17][18][19]. High throughput technology has even been extended to proteomics, where mass spectrometry is being used as a tool in rapid protein identification [20][21][22].Electrospray ionization (ESI), atmospheric pressure chemical ionization (APCI), and matrix-assisted laser desorption/ionization (MALDI) mass spectrometry have become useful for qualitative, and more recently, quantitative analysis, aiding the development of mass spectrometry as a high throughput tool in these different fields (Fig. 1) [2][3][4][9][10][11][12][13][14][15][16].…”