Displacers improve the selectivity of phosphopeptide enrichment by metal oxide affinity chromatography

Yesenia Herrera, Sandra Contreras, Magdalena Hernández, Laura Álvarez, Yolanda Mora, Sergio Encarnación-Guevara
2017 Boletín Médico del Hospital Infantil de México  
KEYWORDS Gram-negative bacteria; Metal oxide affinity chromatography (MOAC); Mass spectrometry; Matrix-assisted laser desorptionionization-time of flight (MALDI-TOF). Abstract Background: A key process in cell regulation is protein phosphorylation, which is catalyzed by protein kinases and phosphatases. However, phosphoproteomics studies are difficult because of the complexity of protein phosphorylation and the number of phosphorylation sites. Methods: We describe an efficient approach
more » ... phosphopeptides in single, separated protein by two-dimensional gel electrophoresis. In this method, a titanium oxide (TiO 2 )-packed NuTip is used as a phosphopeptide trap, together with displacers as lactic acid in the loading buffer to increase the efficiency of the interaction between TiO 2 and phosphorylated peptides. Results: The results were obtained from the comparison of mass spectra of proteolytic peptides of proteins with a matrix-assisted laser desorption-ionization-time of flight (MALDI-TOF) instrument. Conclusions: This method has been applied to identifying phosphoproteins involved in the symbiosis Rhizobium etli-Phaseolus vulgaris.
doi:10.1016/j.bmhimx.2017.03.008 pmid:29382487 fatcat:bodngnojy5bbrjcr3n6hed6lx4