Quantification of intractable membrane proteins in genetically engineered crops by liquid chromatography coupled with tandem mass spectrometry

2017 ◽  
Vol 9 (19) ◽  
pp. 2821-2829 ◽  
Author(s):  
Lindsey J. Schacherer ◽  
Michaela A. Owens ◽  
Tiger X. Hu
Keyword(s):  
Mass Spectrometry ◽  
Liquid Chromatography ◽  
Membrane Proteins ◽  
Membrane Protein ◽  
Tandem Mass Spectrometry ◽  
Genetically Engineered ◽  
Soybean Seeds ◽  
Tandem Mass ◽  
Maize Leaves ◽  
Genetically Engineered Crops

Liquid chromatography with tandem mass spectrometry (LC-MS/MS) methods to quantify a membrane protein in genetically engineered maize leaves and another in soybean seeds were developed and validated.

Analysis of Brassinosteroids in Soybean Seeds and Leaves by Liquid Chromatography-Tandem Mass Spectrometry

2017 ◽  
Vol 10 (1) ◽  
pp. 100-109
Author(s):  
Hongxia Li ◽  
Natasha Trzaskalski ◽  
R.J. Neil Emery
Keyword(s):  
Mass Spectrometry ◽  
Liquid Chromatography ◽  
Tandem Mass Spectrometry ◽  
High Performance ◽  
Soybean Seeds ◽  
Tandem Mass ◽  
Electrospray Tandem Mass Spectrometry ◽  
Aqueous Methanol ◽  
Chromatography Tandem Mass Spectrometry ◽  
Liquid Chromatography Tandem Mass

Objective:A simple and fast high performance liquid chromatography-electrospray tandem mass spectrometry (LC-ESI-MS/MS) method has been developed for the analysis of brassinosteroids (BRs) in plants without derivatization.Materials:The BRs (including castasterone, 24-epicastasterone, brassinolide and 24-epibrassinolide) have been extracted with ice cold 80% aqueous methanol solution.Method:Five different purification strategies have been tested for the purification and enrichment of BRs.Conclusion:This analytical method was sensitive, reliable, rapid and applicable to trace analysis in complex plant samples.

scholarly journals Total, Membrane, and Immunogenic Proteomes of Macrophage- and Tick Cell-Derived Ehrlichia chaffeensis Evaluated by Liquid Chromatography-Tandem Mass Spectrometry and MALDI-TOF Methods

2008 ◽  
Vol 76 (11) ◽  
pp. 4823-4832 ◽  
Author(s):  
Gwi-Moon Seo ◽  
Chuanmin Cheng ◽  
John Tomich ◽  
Roman R. Ganta
Keyword(s):  
Mass Spectrometry ◽  
Liquid Chromatography ◽  
Membrane Proteins ◽  
Tandem Mass Spectrometry ◽  
Tandem Mass ◽  
Tick Cell ◽  
Chromatography Tandem Mass Spectrometry ◽  
Liquid Chromatography Tandem Mass ◽  
Immunogenic Proteins ◽  
Total Membrane

ABSTRACT Ehrlichia chaffeensis, a tick-transmitted rickettsial, is the causative agent of human monocytic ehrlichiosis. To examine protein expression patterns, we analyzed total, membrane, and immunogenic proteomes of E. chaffeensis originating from macrophage and tick cell cultures. Total proteins resolved by one-dimensional gel electrophoresis and subjected to liquid chromatography-electrospray ionization ion trap mass spectrometry allowed identification of 134 and 116 proteins from macrophage- and tick cell-derived E. chaffeensis, respectively. Because a majority of immunogenic proteins remained in the membrane fraction, individually picked total and immunogenic membrane proteins were also surveyed by liquid chromatography-tandem mass spectrometry and matrix-assisted laser desorption ionization-time of flight methods. The analysis aided the identification of 48 additional proteins. In all, 278 genes of the E. chaffeensis genome were verified as functional genes. They included genes for DNA and protein metabolism, energy metabolism and transport, membrane proteins, hypothetical proteins, and many novel proteins of unknown function. The data reported in this study suggest that the membrane of E. chaffeensis is very complex, having many expressed proteins. This study represents the first and the most comprehensive analysis of E. chaffeensis-expressed proteins. This also is the first study confirming the expression of nearly one-fourth of all predicted genes of the E. chaffeensis genome, validating that they are functionally active genes, and demonstrating that classic shotgun proteomic approaches are feasible for tick-transmitted intraphagosomal bacteria. The identity of novel expressed proteins reported in this study, including the large selection of membrane and immunogenic proteins, will be valuable in elucidating pathogenic mechanisms and developing effective prevention and control methods.

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