Human PGP-ATPase Assay
Summary
Summary | Materials
| Solutions | Assay
Procedure |
Human MDR1 (PGP)
P-glycoprotein (PGP), the product of the MDR1 gene, is a member of the
ABC transporter superfamily. PGP is expressed in the
human intestine, liver, kidney and other tissues. Localized in the cell
membrane, PGP functions as an ATP-dependent efflux pump, transporting
many structually unrelated xenobiotics out of cells. Intestinal expression
of PGP may affect the oral bioavailability of drug molecules that are
substrates for this transporter.
The human PGP-ATPase assay is a valuable screening tool for determining
if a drug interacts with PGP. The ATPase assay provides a rapid, colorimetric,
compound-independent measure of the concentration-dependence of any interaction
of a drug with PGP. If a drug does not stimulate PGP-ATPase activity,
the assay can still determine if the drug interacts with PGP as an inhibitor
of the ATPase activity stimulated by a known substrate, such as verapamil.
This technical bulletin describes two step-by-step procedures
for the Human PGP-ATPase assay. The first procedure is for performing
the method in control and/or PGP membranes with 0 or 20 uM PGP substrate,
verapamil. Many users start with this procedure to evaluate the method
in their laboratories. The second procedure describes a drug concentration-dependence
assay in the PGP membranes.
BD Gentest also performs this assay for clients as
one of our contract
research services.
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