Comprehensive identification of erythrocyte membrane protein deficiency by 2D gel electrophoresis based proteomic analysis in hereditary elliptocytosis and spherocytosis.
OBJECTIVE
Sodium dodecyl sulphate-polyacrylamide gel electrophoresis (1DE) may reveal qualitative or quantitative defects in red blood cell (RBC) membrane proteins, two-dimensional gel electrophoresis (2DE) can be used for determination of the protein changes caused by the disease process in a relatively high-throughput manner, because it permits an analysis of thousands of modified or unmodified proteins simultaneously. The principal aim of this study was to compare hereditary elliptocytosis (HE), hereditary spherocytosis (HS), and control RBC membrane protein profiles and identify proteins as a clinical marker by the sensitive methods.


METHODS
RBC membrane proteins of HE, HS, and control groups were compared by 2DE and matrix-assisted laser desorption/ionization-time of flight (MALDI-TOF) MS analysis was obtained using peptide mass fingerprint for protein identification.


RESULTS
Twenty proteins were identified with peptide mass fingerprint analysis and different expression levels were found in HE, HS, and controls for some proteins that includes three biomarker proteins (ankyrin, spectrin, band 3) that may have prognostic or predictive importance.


CONCLUSIONS
2DE of RBC proteins is a potentially valuable method for studying heritable disorders such as HE and HS. By identifying a deficiency in membrane proteins associated with the RBC cytoskeleton, the diagnosis of HE and HS can be confirmed.
Ankara University Biotechnology Institute, Ankara, Turkey. ozel@medicine.ankara.edu.tr
Duygu Ozel
DO
Demiralp
Selen
S
Peker
Burhan
B
Turgut
Nejat
N
Akar
D016022:Case-Control Studies; D015180:Electrophoresis, Gel, Two-Dimensional / Q000379:methods*; D004612:Elliptocytosis, Hereditary / Q000378:metabolism*; D004910:Erythrocyte Membrane / Q000378:metabolism*; D006801:Humans; D008565:Membrane Proteins / Q000172:deficiency* / Q000378:metabolism; D040901:Proteomics / Q000379:methods*; D013103:Spherocytosis, Hereditary / Q000378:metabolism*
D008565:Membrane Proteins