Single molecule tracking of P-glycoprotein in live cells reveals dynamic heterogeneity

P-glycoprotein transports chemotherapy drugs from the plasma membrane and allows cancer cells to survive treatment. We transiently transfected PGP labeled with enhanced green fluorescent protein (PGP-EGFP) into MES-SA cells and used single molecule tracking techniques to characterize the dynamics on the surface of live cells. PGP exhibits freely diffusive behavior at short times and is confined at long times with a transition to anomalous diffusion at 0.7 s.

[1]  F. Lozupone,et al.  P-glycoprotein-actin association through ERM family proteins: a role in P-glycoprotein function in human cells of lymphoid origin. , 2002, Blood.

[2]  R. Marches,et al.  An anti-CD19 antibody inhibits the interaction between P-glycoprotein (P-gp) and CD19, causes P-gp to translocate out of lipid rafts, and chemosensitizes a multidrug-resistant (MDR) lymphoma cell line. , 2004, Blood.

[3]  F. Sharom,et al.  Interaction of the P-glycoprotein multidrug efflux pump with cholesterol: effects on ATPase activity, drug binding and transport. , 2008, Biochemistry.

[4]  Z. Bacso,et al.  Raft and cytoskeleton associations of an ABC transporter: P‐glycoprotein , 2004, Cytometry. Part A : the journal of the International Society for Analytical Cytology.

[5]  Akihiro Kusumi,et al.  Relationship of lipid rafts to transient confinement zones detected by single particle tracking. , 2002, Biophysical journal.

[6]  M. Gottesman,et al.  Multidrug resistance in cancer: role of ATP–dependent transporters , 2002, Nature Reviews Cancer.

[7]  Michelle K. Knowles,et al.  Single secretory granules of live cells recruit syntaxin-1 and synaptosomal associated protein 25 (SNAP-25) in large copy numbers , 2010, Proceedings of the National Academy of Sciences.

[8]  S. Krämer,et al.  Effect of the modulation of the membrane lipid composition on the localization and function of P-glycoprotein in MDR1-MDCK cells , 2005, In Vitro Cellular & Developmental Biology - Animal.

[9]  P. Koumoutsakos,et al.  Feature point tracking and trajectory analysis for video imaging in cell biology. , 2005, Journal of structural biology.

[10]  S. Krämer,et al.  P-Glycoprotein in Proteoliposomes with Low Residual Detergent: The Effects of Cholesterol , 2007, Pharmaceutical Research.

[11]  T. Fojo,et al.  The role of efflux pumps in drug-resistant metastatic breast cancer: new insights and treatment strategies. , 2007, Clinical breast cancer.

[12]  T. Kiss,et al.  P-glycoprotein inhibition by membrane cholesterol modulation. , 2008, European journal of pharmaceutical sciences : official journal of the European Federation for Pharmaceutical Sciences.

[13]  H. Grunicke,et al.  Rapid functional assay for the detection of multidrug-resistant cells using the fluorescent dye rhodamine 123. , 1991, Blood.

[14]  M. Tamkun,et al.  A cytoskeletal-based perimeter fence selectively corrals a sub-population of cell surface Kv2.1 channels , 2007, Journal of Cell Science.

[15]  F. Sharom,et al.  P‐Glycoprotein is localized in intermediate‐density membrane microdomains distinct from classical lipid rafts and caveolar domains , 2005, The FEBS journal.

[16]  V. Ling,et al.  P-glycoprotein expression as a predictor of the outcome of therapy for neuroblastoma. , 1991, The New England journal of medicine.

[17]  H. Omote,et al.  The Remarkable Transport Mechanism of P-Glycoprotein: A Multidrug Transporter , 2005, Journal of bioenergetics and biomembranes.

[18]  J. Troost,et al.  Cholesterol modulates P-glycoprotein activity in human peripheral blood mononuclear cells. , 2004, Biochemical and biophysical research communications.

[19]  I. Pastan,et al.  Biochemical, cellular, and pharmacological aspects of the multidrug transporter. , 1999, Annual review of pharmacology and toxicology.

[20]  D. Grier,et al.  Methods of Digital Video Microscopy for Colloidal Studies , 1996 .

[21]  Ronald D. Vale,et al.  Single-Molecule Microscopy Reveals Plasma Membrane Microdomains Created by Protein-Protein Networks that Exclude or Trap Signaling Molecules in T Cells , 2005, Cell.