A novel electrochemical method for efficient reduction of disulfide bonds in peptides and proteins prior to MS detection

Kraj, Agnieszka; Brouwer, Hendrik-Jan; Reinhoud, Nico; Chervet, Jean-Pierre
November 2013
Analytical & Bioanalytical Chemistry;Nov2013, Vol. 405 Issue 27, p9311
Academic Journal
A novel electrochemical (EC) method for fast and efficient reduction of the disulfide bonds in proteins and peptides is presented. The method does not use any chemical agents and is purely instrumental. To demonstrate the performance of the EC reactor cell online with electrospray mass spectrometry, insulin and somatostatin were used as model compounds. Efficient reduction is achieved in continuous infusion mode using an EC reactor cell with a titanium-based working electrode. Under optimized conditions, the presented method shows almost complete reduction of insulin and somatostatin. The method does not require any special sample preparation, and the EC reactor cell makes it suitable for automation. Online EC reduction followed by collision-induced dissociation fragmentation of somatostatin showed more backbone cleavages and improved sequence coverage. By adjusting the settings, the EC reaction efficiency was gradually changed from partial to full disulfide bonds reduction in α-lactalbumin, and the expected shift in charge state distribution has been demonstrated. The reduction can be controlled by adjusting the square-wave pulse, flow rate or mobile phase composition. We have shown the successful use of an EC reactor cell for fast and efficient reduction of disulfide bonds for online mass spectrometry of proteins and peptides. The possibility of online and gradual disulfide bond reduction adds a unique dimension to characterization of disulfide bonds in mid- and top-down proteomics applications. [Figure not available: see fulltext.]


Related Articles

  • Recent Advances and Special Considerations for the Analysis of Phosphorylated Peptides by LC-ESI-MS/MS. Suya Liu; Hughes, Chris; Lajoie, Gilles A. // Current Analytical Chemistry;Jan2012, Vol. 8 Issue 1, p35 

    Liquid chromatography electrospray tandem mass spectrometry (LC-ESI-MS/MS) is the primary method for sample analysis used in proteomics. The high-resolution separation of liquid chromatography (LC), coupled with the sensitivity and specificity of mass spectrometry (MS) permits the identification...

  • A Survey of Membrane Proteins in Human Serum. Nguyen Tien Dung; Phan Van Chi // Proteomics Insights;2012, Issue 5, p1 

    Serum and membrane proteins are two of the most attractive targets for proteomic analysis. Previous membrane protein studies tend to focus on tissue sample, while membrane protein studies in serum are still limited. In this study, an analysis of membrane proteins in normal human serum was...

  • Prospects for monolithic nano-LC columns in shotgun proteomics. Guryča, Vilém; Kieffer-Jaquinod, Sylvie; Garin, Jerôme; Masselon, Christophe // Analytical & Bioanalytical Chemistry;Dec2008, Vol. 392 Issue 7/8, p1291 

    We report a premier side-by-side comparison of two leading types of monolithic nano-LC column (silica-C18, polystyrene) in shotgun proteomics experiments. Besides comparing the columns in terms of the number of peptides from a real-life sample ( Arabidopsis thaliana chloroplast) that they...

  • Sample Preparation Guide for Mass Spectrometry-Based Proteomics. Hess, Sonja // Spectroscopy;Jul2013 Current Trends in Mass Spectrometry, p12 

    The article presents successful sample preparation strategies for mass spectrometry (MS)-based proteomics experiment. It discusses good sample preparation techniques depending on the experiment technique used and biological sample selected. It emphasizes on preparation of a proper experimental...

  • Derivatization for Ultratrace Quantitative Analysis Using HPLC-ESI-MS. Schug, Kevin A. // LC-GC North America;Apr2013, Vol. 31 Issue 4, p334 

    A blog related to the derivatization technique electrospray ionization (ESI) is presented.

  • Erratum to: Mechanical ion gate for electrospray-ionization ion-mobility spectrometry. Li Zhou; Collins, David C.; Lee, Edgar D.; Rockwood, Alan L.; Lee, Milton L. // Analytical & Bioanalytical Chemistry;Oct2010, Vol. 398 Issue 4, p1833 

    A correction to the article "Mechanical Ion Gate for Electrospray-Ionization Ion-Mobility Spectrometry" that was published in the online version of the journal on August 14, 2010 issue is presented.

  • Mass analysis of receptor?DNA complexes. DeWitt, Natalie // Nature Biotechnology;Dec99, Vol. 17 Issue 12, p1150 

    Focuses on the use of electrospray ionization mass spectrometry (ESI-MS) in the study of protein complexes. Use of ESI-MS in the analysis of the effect of ligands on heterodimerization of full-length vitamin D and retinoid X receptors.

  • Comparison of Dissociation of Ions in an Electrospray Source, or a Collision Cell in Tandem Mass Spectrometry. Bure, Corinne; Lange, Catherine // Current Organic Chemistry;Oct2003, Vol. 7 Issue 15, p1613 

    Electrospray ionization (ESI) is a soft ionization technique well suited for producing gas phase ions of biological macromolecules. The use of ESI when combined with collision-induced dissociation and mass analysis can give structural information about biomolecules. Traditionally, collisional...

  • Ion Traps Provide High-Throughput Analysis. Stafford, George // R&D Magazine;Nov2001, Vol. 43 Issue 11, p49 

    Presents information on the unique properties of quadrupole ion trap mass spectrometry (MS) that make it a versatile technology for researchers in both chemistry and biology. Features of the ion trap MS device called LCQ Deca XP from Thermo Finnigan company; Difference of quadrupoles from ions...


Read the Article


Sorry, but this item is not currently available from your library.

Try another library?
Sign out of this library

Other Topics