Optimization of Western blotting analysis for the isolation and detection of membrane xenobiotic transporter ABCG2
| Autor: | Małgorzata Szczygieł, Roxana Zuziak, Milena Julia Szafraniec, Marcin Markiewicz, Krystyna Urbanska, Leszek Fiedor |
|---|---|
| Rok vydání: | 2017 |
| Předmět: |
Male
0301 basic medicine xenobiotic transporter Abcg2 ABCG2 Blotting Western Detergents Kidney General Biochemistry Genetics and Molecular Biology Western blotting Xenobiotics Epitopes 03 medical and health sciences chemistry.chemical_compound ATP hydrolysis ATP Binding Cassette Transporter Subfamily G Member 2 Animals Integral membrane protein biology Transporter biology.organism_classification Immunohistochemistry Blot Transmembrane domain 030104 developmental biology Biochemistry chemistry Mice Inbred DBA biology.protein BCRP Xenobiotic Bacteria |
| Zdroj: | Acta Biochimica Polonica. 64:437-443 |
| ISSN: | 1734-154X 0001-527X |
| DOI: | 10.18388/abp.2017_2299 |
| Popis: | All organisms are exposed to numerous stress factors, which include harmful xenobiotics. The diversity of these compounds is enormous, thus in the course of evolution diverse biological defense mechanisms at various levels of organization have developed. One of them engages an evolutionarily conserved family of transporters from the ABC superfamily, found in most species - from bacteria to humans. An important example of such a transporter is the breast cancer resistance protein (BCRP/ABCG2), a typical integral membrane protein. It plays a key role in the absorption, distribution and elimination of a wide variety of xenobiotics, including drugs used in chemotherapy, and is involved in multidrug resistance. It also protects against phototoxic chlorophyll derivatives of dietary origin. BCRP is a hemitransporter which consists of one transmembrane domain, made of six alpha-helices forming a characteristic pore structure, and one ATP-binding domain, which provides the energy from ATP hydrolysis, required for active transport of the substrates. The isolation of BCRP is still not an easy task, because its insolubility in water and the presence of membrane rafts pose serious methodological and technical challenges during the purification. The aim of this study was to optimize the methods for detection and isolation of BCRP-enriched fractions obtained from animal tissue samples. In this report we describe an optimization of isolation of a BCRP-enriched membrane fraction, which is suitable for further protein quantitative and qualitative analysis using the molecular biology tools. |
| Databáze: | OpenAIRE |
| Externí odkaz: |
|