Description
General description
Membrane Preparations for Vesicular Transport Assays (VT) are suitable for general drug-efflux transporter interaction studies. Both substrate and inhibitor interactions can be assessed using vesicles. The success of substrate interaction studies strongly depends on the passive permeability of the compound. High permeability substrates might not be detected. Control Membranes with no-, or significantly lower transporter activity are also available.
Multidrug resistance associated protein 4 (MRP4), also known as ATP binding cassette subfamily C member 4 (ABCC4), is encoded by the gene mapped to human chromosome 13q32.1. The encoded protein belongs to the family of ATP-binding cassette transporters involved in ATP driven transmembranous transport of substrates.
Application
In the vesicular transport assay so-called "inside-out" membrane vesicles containing ABC transporters are applied. Incubating substrates of the respective efflux transporter in the presence of the inverted membrane vesicles and ATP will allow measuring accumulation of the substrates into the vesicles. In many cases radiolabeled reporter substrates are used but recently SOLVO developed the new PREDIVEZTM Vesicular Transport Kits that use fluorescent reporter substrates.
The standard vesicular transport assay is an inhibitory assay performed with cold test articles. This assay provides information on any interaction between the ABC transporter and the test article. The transport of the reporter substrate is measured in the presence of the test article (typically in 7 concentrations) and IC50 is defined as the concentration inhibiting the transport of the reporter substrate by 50%.
Should radiolabeled form of the investigated compound or adequate analytical methods (LC/MS, HPLC) be available, the vesicular transport assay may be performed in a direct format without the reporter substrate and may identify substrate nature of the test article. The vesicular transport substrate assay is a low throughput assay. It is suitable for low permeability test compounds as high permeability compounds may escape from the vesicles through the lipid bilayer.
SB-MRP4-HEK293 has been used in vesicular transport study.
Biochem/physiol Actions
Multidrug resistance associated protein 4 (MRP4) plays a vital role in substrate-specific transport of endogenous and exogenous substrates. It also has a role in non-neoplastic conditions. MRP4 also contributes to bile secretion and various other physiological roles. Upregulated expression of the gene is associated with the pancreatic cancer development. Thus, MRP4 can be considered as a potent therapeutic target for pancreatic cancer.
Physical form
Supplied as frozen membrane vesicles, containing 5 mg/ml membrane protein, labeled with volume, catalog number (transporter) and date of production.
Legal Information
Distributed for SOLVO Biotechnology, Inc.
