The recent x-ray structure of mouse P-glycoprotein (Pgp) determined to 3.8 angstrom established the overall structural architecture of one of the most studied mammalian multidrug resistance (MDR) transporters. The structure of Pgp, however, is only a starting point for understanding its transport mechanism and potential inhibition. Clearly a higher resolution structure along with different conformations and co-crystal structures will be required to understand the detailed mechanism of transport coupled to ATP hydrolysis paving the way towards future rational drug design. We propose to (1) extend the resolution of Pgp crystals using very new membrane protein crystallization techniques and novel detergents to greatly improve the precision of the model, (2) determine additional conformations of Pgp both in the inward- and outward- facing conformations to map out the structural trajectories of the transport cycle, (3) determine the co-crystal structures of Pgp of two classical and clinically important compounds, doxorubicin and rhodamine, to understand poly-specific substrate binding, and (4) determine the x-ray structure of human Pgp or humanized version of Pgp, which will be very useful for drug design in the future.

Public Health Relevance

P-glycoprotein plays a significant role in drug efficacy and multidrug resistance to several diseases, including cancer and HIV. This high-impact project will provide a detailed molecular structural basis underlying its transport mechanism. The structures of Pgp should greatly facilitate the development of new drugs to circumvent multidrug resistance in the future.

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Project (R01)
Project #
5R01GM094367-05
Application #
8536320
Study Section
Biochemistry and Biophysics of Membranes Study Section (BBM)
Program Officer
Chin, Jean
Project Start
2010-09-20
Project End
2014-08-31
Budget Start
2013-09-01
Budget End
2014-08-31
Support Year
5
Fiscal Year
2013
Total Cost
$284,667
Indirect Cost
$72,974
Name
University of California San Diego
Department
Pharmacology
Type
Schools of Pharmacy
DUNS #
804355790
City
La Jolla
State
CA
Country
United States
Zip Code
92093
Hill, Jacqueline E; Linder, Michelle K; Davies, Kellie S et al. (2014) Selenorhodamine photosensitizers for photodynamic therapy of P-glycoprotein-expressing cancer cells. J Med Chem 57:8622-34
Martinez, Lorena; Arnaud, Ophelie; Henin, Emilie et al. (2014) Understanding polyspecificity within the substrate-binding cavity of the human multidrug resistance P-glycoprotein. FEBS J 281:673-82
Ward, Andrew B; Szewczyk, Paul; Grimard, Vinciane et al. (2013) Structures of P-glycoprotein reveal its conformational flexibility and an epitope on the nucleotide-binding domain. Proc Natl Acad Sci U S A 110:13386-91
Myette, Robert L; Conseil, Gwenaelle; Ebert, Sean P et al. (2013) Chalcogenopyrylium dyes as differential modulators of organic anion transport by multidrug resistance protein 1 (MRP1), MRP2, and MRP4. Drug Metab Dispos 41:1231-9
Ebert, Sean P; Wetzel, Bryan; Myette, Robert L et al. (2012) Chalcogenopyrylium compounds as modulators of the ATP-binding cassette transporters P-glycoprotein (P-gp/ABCB1) and multidrug resistance protein 1 (MRP1/ABCC1). J Med Chem 55:4683-99