Abstract
We have developed a rapid, simple and reliable, antibody-based flow cytometry assay for the quantitative determination of membrane proteins in human erythrocytes. Our method reveals significant differences between the expression levels of the wild-type ABCG2 protein and the heterozygous Q141K polymorphic variant. Moreover, we find that nonsense mutations on one allele result in a 50% reduction in the erythrocyte expression of this protein. Since ABCG2 polymorphisms are known to modify essential pharmacokinetic parameters, uric acid metabolism and cancer drug resistance, a direct determination of the erythrocyte membrane ABCG2 protein expression may provide valuable information for assessing these conditions or for devising drug treatments. Our findings suggest that erythrocyte membrane protein levels may reflect genotype-dependent tissue expression patterns. Extension of this methodology to other disease-related or pharmacologically important membrane proteins may yield new protein biomarkers for personalized diagnostics.
Publication types
-
Research Support, Non-U.S. Gov't
MeSH terms
-
ATP Binding Cassette Transporter, Subfamily G, Member 2
-
ATP-Binding Cassette Transporters / genetics
-
ATP-Binding Cassette Transporters / metabolism*
-
Codon, Nonsense / genetics
-
Erythrocytes / metabolism*
-
Flow Cytometry / methods*
-
Gene Expression / genetics*
-
Humans
-
Mutation, Missense / genetics
-
Neoplasm Proteins / genetics
-
Neoplasm Proteins / metabolism*
Substances
-
ABCG2 protein, human
-
ATP Binding Cassette Transporter, Subfamily G, Member 2
-
ATP-Binding Cassette Transporters
-
Codon, Nonsense
-
Neoplasm Proteins
Grants and funding
This work has been supported by grants from OTKA (Hungarian: National Scientific Research Fund) NK83533 and KMOP (The Central Hungary Region Operational Programme) –1.1.2–07/1–2008–0003. Gergely Szakács is supported by the Lendület grant from the Hungarian Academy of Sciences. Hajnalka Andrikovics is a recipient of the Janos Bolyai Research Scholarship from the Hungarian Academy of Sciences. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.