Mass spectrometry protein analysis

Liang, Z., Duan, J., Zhang, L., Zhang, W., Zhang, Y., and Yan, C. (2004). Pressurized electrochromatography coupled with electrospray ionization mass spectrometry for analysis of peptides and proteins. Anal. Chem. 76, 6935-6940. 

The practice of protein analysis of whole proteomes relies on (i) two-dimensional gel electrophoresis for separation (ii) mass spectrometry for analysis and (iii) protein arrays for achieving massively parallel analysis. 

Protein Sequencing Mass Spectrometry Proteins were sequenced on an Applied Biosystems 477A protein sequencer. Polypeptide molecular weights were determined after analysis by a Sciex API-Ill electrospray ioniziation mass spectrometer. 

Medzihradszky, K.F., H. Leffler, M.A. Baldwin and A. Burlinghame. Protein identification by in-gel digestion, high-performance liquid chromatography and mass spectrometry peptide analysis by complementary ionization techniques. J. Am. Soc. Mass Spectrom. 12 215-221, 2001. 

Sanz-Nebot V, Benavente F, Gimenez E, et al. (2005). Capillary electrophoresis and matrix-assisted laser desorption/ionization-time of flight-mass spectrometry for analysis of the novel erythropoiesis-stimulating protein (NESP). Electrophoresis. 26 1451-1456. 

Myung, S., Wiseman, J.M., Valentine, S.J., Takats, Z., Cooks, R.G., Clemmer, D.E., Coupling desorption electrospray ionization with ion mohihty/mass spectrometry for analysis of protein structure evidence for desorption of folded and denatured states. J. Phys. Chem. B 2006,110, 5045. 

Figure 1.2 The classical continuous labeling (bottom-up) HX-MS experiment. HX of an equilibrated protein solution is initiated by dilution into a Dfl-containing buffer, and exchange is quenched at various time points. Global HX (protein level) can be measured directly by liquid chromatography (LC) and mass spectrometry (MS) analysis of the intact protein, or local HX (peptide level) can be measured by enzymatic cleavage and subsequent LC-MS analysis of the proteolytic peptides. (See insert for color representation of the figure.)...

Sharma, S., Zheng, H., Huang, Y.J., et al. (2009) Construct optimization for protein NMR structure analysis using amide hydrogen/deuterium exchange mass spectrometry. Proteins, 76 (4), 882-894. 

Schmidt A-C, Fahlbusch B, Otto M. Size exclusion chromatography coupled to electrospray ionization mass spectrometry for analysis and quantitative characterization of arsenic interactions with peptides and proteins. J Mass Spectrom 2009 44 898-910. 

The first reports of metabolic labeling appeared in 1999. In this procedure, yeast cells were grown in two media, one of which used N-emiehed media. " The two yeast cultures were combined and the proteins of interest were digested with trypsin before mass speetrometry analysis. The labeling by use of an N-ammonium salt results in the eomplete labeling of amino acid and could be quantified with mass spectrometry. The corresponding mass shift between the unlabeled and the labeled form of the peptide requires high resolution mass spectrometry for analysis. 

Wen DX, Livingston BD, Medzihradszky KF, Kelm S, Burlingame AL, et al. Primary Structure of Galbetal,3(4)GlcNAc 2,3-Sialyltransferase Determined by Mass Spectrometry Sequence Analysis and Molecular Cloning—Evidence for a Protein Motif in the Sialyltransferase Gene Family. 7. Biol. Chem. 1992 267 21011 21019. 

Until 1981, mass spectrometry was limited, generally, to the analysis of volatile, relatively low-molecular-mass samples and was difficult to apply to nonvolatile peptides and proteins without first cutting them chemically into smaller volatile segments. During the past decade, the situation has changed radically with the advent of new ionization techniques and the development of tandem mass spectrometry. Now, the mass spectrometer has a well-deserved place in any laboratory interested in the analysis of peptides and proteins. 

The techniques described thus far cope well with samples up to 10 kDa. Molecular mass determinations on peptides can be used to identify modifications occurring after the protein has been assembled according to its DNA code (post-translation), to map a protein structure, or simply to confirm the composition of a peptide. For samples with molecular masses in excess of 10 kDa, the sensitivity of FAB is quite low, and such analyses are far from routine. Two new developments have extended the scope of mass spectrometry even further to the analysis of peptides and proteins of high mass. 

Chapter 40 Analysis of Peptides and Proteins by Mass Spectrometry... 

The use of mass spectrometry for the analysis of peptides, proteins, and enzymes has been summarized. This chapter should be read in conjunction with others, including Chapter 45, An Introduction to Biotechnology, and Chapters 1 through 5, which describe specific ionization techniques in detail. 

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