Protein profiling of sickle cell versus control RBC core membrane skeletons by ICAT technology and tandem mass spectrometry.
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 (alpha spectrin, beta 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.
Chou
Jose
J
Department of Molecular and Cell Biology, University of Texas at Dallas, Richardson, Texas, USA.
Choudhary
Pankaj K
PK
Goodman
Steven R
SR
D000755:Anemia, Sickle Cell / Q000473:pathology*; D016022:Case-Control Studies; D016001:Confidence Intervals; D004910:Erythrocyte Membrane / Q000737:chemistry*; D004913:Erythrocytes, Abnormal / Q000737:chemistry*; D006801:Humans; D007553:Isotope Labeling; D008565:Membrane Proteins / Q000032:analysis / Q000737:chemistry*; D040901:Proteomics; D053719:Tandem Mass Spectrometry
D008565:Membrane Proteins