Protein profiling of sickle cell versus control RBC core membrane skeletons by ICAT technology and tandem mass spectrometry
Cellular & Molecular Biology Letters volume 11, pages 326–337 (2006)
A proteomic approach using a cleavable ICAT reagent and nano-LC ESI tandem mass spectrometry was used to perform protein profiling of core RBC membrane skeleton proteins between sickle cell patients (SS) and controls (AA), and determine the efficacy of this technology. The data was validated through Peptide/Protein Prophet and protein ratios were calculated through ASAPratio. Through an ANOVA test, it was determined that there is no significant difference in the mean ratios from control populations (AA1/AA2) and sickle cell versus control populations (AA/SS). The mean ratios were not significantly different from 1.0 in either comparison for the core skeleton proteins (α spectrin, β spectrin, band 4.1 and actin). On the natural-log scale, the variation (standard deviation) of the method was determined to be 14.1% and the variation contributed by the samples was 13.8% which together give a total variation of 19.7% in the ratios.
automated statistical analysis of protein abundance ratios+
cleavable isotope coded affinity tag
collision induced dissociation
- 2D DIGE:
two dimension differential gel electrophoresis
electrospray ionization source
isotope coded affinity tag
red blood cell
stable isotope labeled amino acids in cell culture
white blood cells
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Chou, J., Choudhary, P.K. & Goodman, S.R. Protein profiling of sickle cell versus control RBC core membrane skeletons by ICAT technology and tandem mass spectrometry. Cell Mol Biol Lett 11, 326–337 (2006). https://doi.org/10.2478/s11658-006-0026-2
- Cleavable ICAT
- Ion trap mass spectrometry
- RBC membrane skeleton
- Sickle cell