European Biophysics Journal

, Volume 36, Issue 2, pp 121–131 | Cite as

Multi drug resistance-dependent “vacuum cleaner” functionality potentially driven by the interactions between endocytosis, drug size and Pgp-like transporters surface density



In cells, multi drug resistance (MDR) is associated with Pgp-like transporters expression extruding drugs from cellular membranes. MDR is efficiently generated with a relatively small fraction of membrane transporters. As the insertion of drugs into cellular membranes is widespread, there are no reasons why a drug should incorporate the membrane in the vicinity of a transporter. As a result a further elusive hypothesis is usually invoked: these transporters act like “vacuum cleaners” of drugs embedded in the membrane. Nonetheless, how these transporters attract drugs remains obscure. To clarify the “vacuum cleaner” notion, we suggest that during its residency time in cellular membranes, the lateral movement of drugs from their point of insertion to transporters is governed by Brownian’s diffusion, which allows the drugs/transporters interaction. Taking into account the functionality of Pgp-like transporters, namely the extrusion of drugs from the plasma membrane inner leaflet, we characterize how the state of drug resistance is triggered involving: membrane endocytosis, drug physico-chemical properties and the surface density of Pgp-like transporters. In addition, the theory developed provides for the first time a theoretical proof of Lipinski’s second rule with regard to drugs’ size (or MW) selectivity on their permeation across cellular membranes.

List of symbols


drug cross section area


membrane diffusion coefficient


non-recurring step number of a two dimensional random walk


drug dehydration energy


membrane thickness


bending modulus of the membrane

k or k0

altered or control kinetics of endocytosis


step number of a two dimensional random walk


number of Pgps in the outer cellular surface


meeting probability between a drug and a Pgp

\({ \tilde{p}_{{\rm Pgp}}}\)

drug extrusion probability by Pgp

rMDR, rnon-MDR

escape rate (i.e. probability per unit of time) into the cytoplasm of drugs in the membrane of drug resistant (“MDR”) and drug sensitive (“non-MDR”) cells


vesicle radius


cellular surface area


cross section area of Pgps in the cellular surface


drug residency time in the membrane


membrane barrier potential


fraction of the cellular surface covered by Pgp transporters


critical surface area covered by Pgp-like transporters leading to drug resistance


inner leaflet surface tension


outer and inner leaflet surface tension

Δσ = σin − σout

difference of surface tension between the inner and outer leaflets

χMDR, χnon-MDR

ratio between the endocytosis kinetics and the escape rate into the cytoplasm of drugs initially in the plasma membrane of resistant (“MDR”) and sensitive (“non-MDR”) cells



The authors are grateful to Dr Emmanuel Farge and Zoe Rauch for their comments on the manuscript. This work has been supported by BBSRC (Biotechnology and Biological Sciences Research Council, UK), Grant Nos: BB/C505308/1.

Supplementary material


  1. Altan N, Chen Y, Schindler M, Simon SM (1998) Defective acidification in human breast tumor cells and implications for chemotherapy. J Exp Med 187(10):1583–1598CrossRefGoogle Scholar
  2. Altan N, Chen Y, Schindler M, Simon SM (1999) Tamoxifen inhibits acidification in cells independent of the estrogen receptor. Proc Natl Acad Sci USA 96(8):4432–4437CrossRefADSGoogle Scholar
  3. Ambudkar SV, Dey S, Hrycyna CA, Ramachandra M, Pastan I, Gottesman MM (1999) Biochemical, cellular, and pharmacological aspects of the multidrug transporter. Annu Rev Pharmacol Toxicol 39:361–398CrossRefGoogle Scholar
  4. Bloom M, Evans E, Mouritsen OG (1991) Physical properties of the fluid lipid-bilayer component of cell membranes: a perspective. Q Rev Biophys 24(3):293–397Google Scholar
  5. Bornmann WG, Roepe PD (1994) Analysis of drug transport kinetics in multidrug-resistant cells using a novel coumarin–vinblastine compound. Biochemistry 33(42):12665–12675CrossRefGoogle Scholar
  6. Callaghan R, Stafford A, Epand RM (1993) Increased accumulation of drugs in a multidrug resistant cell line by alteration of membrane biophysical properties. Biochim Biophys Acta 1175(3):277–282CrossRefGoogle Scholar
  7. Cano-Gauci DF, Riordan JR (1987) Action of calcium antagonists on multidrug resistant cells. Specific cytotoxicity independent of increased cancer drug accumulation. Biochem Pharmacol 36(13):2115–2123CrossRefGoogle Scholar
  8. Cantor RS (1999) Lipid composition and the lateral pressure profile in bilayers. Biophys J 76(5):2625–2639Google Scholar
  9. Cass CE, Janowska-Wieczorek A, Lynch MA, Sheinin H, Hindenburg AA, Beck WT (1989) Effect of duration of exposure to verapamil on vincristine activity against multidrug-resistant human leukemic cell lines. Cancer Res 49(21):5798–5804Google Scholar
  10. Chan LM, Lowes S, Hirst BH (2004) The ABCs of drug transport in intestine and liver: efflux proteins limiting drug absorption and bioavailability. Eur J Pharm Sci 21(1):25–51CrossRefGoogle Scholar
  11. Cirilli M, Bachechi F, Ughetto G, Colonna FP, Capobianco ML (1993) Interactions between morpholinyl anthracyclines and DNA. The crystal structure of a morpholino doxorubicin bound to d(CGTACG). J Mol Biol 230(3):878–889CrossRefGoogle Scholar
  12. Davoust J, Gruenberg J, Howell KE (1987) Two threshold values of low pH block endocytosis at different stages. Embo J 6(12):3601–3609Google Scholar
  13. Devaux PF, Zachowski A, Favre E, Fellmann P, Cribier S, Geldwerth D, Herve P, Seigneuret M (1986) Energy-dependent translocation of amino-phospholipids in the erythrocyte membrane. Biochimie 68(3):383–393CrossRefGoogle Scholar
  14. Drori S, Eytan GD, Assaraf YG (1995) Potentiation of anticancer-drug cytotoxicity by multidrug-resistance chemosensitizers involves alterations in membrane fluidity leading to increased membrane permeability. Eur J Biochem 228(3):1020–1029CrossRefGoogle Scholar
  15. Dudeja PK, Anderson KM, Harris JS, Buckingham L, Coon JS (1995) Reversal of multidrug resistance phenotype by surfactants: relationship to membrane lipid fluidity. Arch Biochem Biophys 319(1):309–315CrossRefGoogle Scholar
  16. Eytan GD (2005) Mechanism of multidrug resistance in relation to passive membrane permeation. Biomed Pharmacother 59(3):90–97CrossRefGoogle Scholar
  17. Farge E (1995) Increased vesicle endocytosis due to an increase in the plasma membrane phosphatidylserine concentration. Biophys J 69(6):2501–2506Google Scholar
  18. Farge E, Ojcius DM, Subtil A, Dautry-Varsat A (1999) Enhancement of endocytosis due to aminophospholipid transport across the plasma membrane of living cells. Am J Physiol 276(3 Pt 1):C725–733Google Scholar
  19. Ferte J (2000) Analysis of the tangled relationships between P-glycoprotein-mediated multidrug resistance and the lipid phase of the cell membrane. Eur J Biochem 267(2):277–294CrossRefGoogle Scholar
  20. Frezard F, Garnier-Suillerot A (1998) Permeability of lipid bilayer to anthracycline derivatives. Role of the bilayer composition and of the temperature. Biochim Biophys Acta 1389(1):13–22Google Scholar
  21. Germann UA (1996) P-glycoprotein—a mediator of multidrug resistance in tumour cells. Eur J Cancer 32A(6):927–944CrossRefGoogle Scholar
  22. Harguindey S, Orive G, Luis Pedraz J, Paradiso A, Reshkin SJ (2005) The role of pH dynamics and the Na+/H+ antiporter in the etiopathogenesis and treatment of cancer. Two faces of the same coin—one single nature. Biochim Biophys Acta 1756(1):1–24Google Scholar
  23. Heijn M, Roberge S, Jain RK (1999) Cellular membrane permeability of anthracyclines does not correlate with their delivery in a tissue-isolated tumor. Cancer Res 59(17):4458–4463Google Scholar
  24. Heywang C, Saint-Pierre Chazalet M, Masson CM, Bolard J (1998) Orientation of anthracyclines in lipid monolayers and planar asymmetrical bilayers: a surface-enhanced resonance Raman scattering study. Biophys J 75(5):2368–2381Google Scholar
  25. Horton JK, Thimmaiah KN, Harwood FC, Kuttesch JF, Houghton PJ (1993) Pharmacological characterization of N-substituted phenoxazines directed toward reversing Vinca alkaloid resistance in multidrug-resistant cancer cells. Mol Pharmacol 44(3):552–559Google Scholar
  26. Kim H, Barroso M, Samanta R, Greenberger L, Sztul E (1997) Experimentally induced changes in the endocytic traffic of P-glycoprotein alter drug resistance of cancer cells. Am J Physiol 273(2 Pt 1):C687–702Google Scholar
  27. Kivisto KT, Niemi M, Fromm MF (2004) Functional interaction of intestinal CYP3A4 and P-glycoprotein. Fundam Clin Pharmacol 18(6):621–626CrossRefGoogle Scholar
  28. Lee JY, Urbatsch IL, Senior AE, Wilkens S (2002) Projection structure of P-glycoprotein by electron microscopy. Evidence for a closed conformation of the nucleotide binding domains. J Biol Chem 277(42):40125–40131CrossRefGoogle Scholar
  29. Liang X, Huang Y (2002) Physical state changes of membrane lipids in human lung adenocarcinoma A(549) cells and their resistance to cisplatin. Int J Biochem Cell Biol 34(10):1248–1255CrossRefGoogle Scholar
  30. Lindgren CA, Emery DG, Haydon PG (1997) Intracellular acidification reversibly reduces endocytosis at the neuromuscular junction. J Neurosci 17(9):3074–3084Google Scholar
  31. Lipinski CA, Lombardo F, Dominy BW, Feeney PJ (2001) Experimental and computational approaches to estimate solubility and permeability in drug discovery and development settings. Adv Drug Deliv Rev 46(1–3):3–26CrossRefGoogle Scholar
  32. Mao Q, Scarborough GA (1997) Purification of functional human P-glycoprotein expressed in Saccharomyces cerevisiae. Biochim Biophys Acta 1327(1):107–118CrossRefGoogle Scholar
  33. Mitragotri S, Johnson ME, Blankschtein D, Langer R (1999) An analysis of the size selectivity of solute partitioning, diffusion, and permeation across lipid bilayers. Biophys J 77(3):1268–1283Google Scholar
  34. Nielsen D, Maare C, Skovsgaard T (1995) Influx of daunorubicin in multidrug resistant Ehrlich ascites tumour cells: correlation to expression of P-glycoprotein and efflux. Influence of verapamil. Biochem Pharmacol 50(4):443–450CrossRefGoogle Scholar
  35. Ramu A, Glaubiger D, Magrath IT, Joshi A (1983) Plasma membrane lipid structural order in doxorubicin-sensitive and -resistant P388 cells. Cancer Res 43(11):5533–5537Google Scholar
  36. Ramu A, Pollard HB, Rosario LM (1989) Doxorubicin resistance in P388 leukemia—evidence for reduced drug influx. Int J Cancer 44(3):539–547Google Scholar
  37. Rao VV, Herman LW, Kronauge JF, Piwnica-Worms D (1998) A novel areneisonitrile Tc complex inhibits the transport activity of MDR P-glycoprotein. Nucl Med Biol 25(3):225–232CrossRefGoogle Scholar
  38. Rauch C, Farge E (2000) Endocytosis switch controlled by transmembrane osmotic pressure and phospholipid number asymmetry. Biophys J 78(6):3036–3047Google Scholar
  39. Raucher D, Sheetz MP (1999) Membrane expansion increases endocytosis rate during mitosis. J Cell Biol 144(3):497–506CrossRefGoogle Scholar
  40. Raviv Y, Pollard HB, Bruggemann EP, Pastan I, Gottesman MM (1990) Photosensitized labeling of a functional multidrug transporter in living drug-resistant tumor cells. J Biol Chem 265(7):3975–3980Google Scholar
  41. Regev R, Assaraf YG, Eytan GD (1999) Membrane fluidization by ether, other anesthetics, and certain agents abolishes P-glycoprotein ATPase activity and modulates efflux from multidrug-resistant cells. Eur J Biochem 259(1–2):18–24CrossRefGoogle Scholar
  42. Regev R, Yeheskely-Hayon D, Katzir H, Eytan GD (2005) Transport of anthracyclines and mitoxantrone across membranes by a flip-flop mechanism. Biochem Pharmacol 70(1):161–169CrossRefGoogle Scholar
  43. Rudnick J, Gaspari G (2004) Elements of the Random walk. Cambridge University Press, CambridgeMATHGoogle Scholar
  44. Sandvig K, van Deurs B (1994) Endocytosis without clathrin. Trends Cell Biol 4(8):275–277CrossRefGoogle Scholar
  45. Sandvig K, Olsnes S, Petersen OW, van Deurs B (1987) Acidification of the cytosol inhibits endocytosis from coated pits. J Cell Biol 105(2):679–689CrossRefGoogle Scholar
  46. Sandvig K, Olsnes S, Petersen OW, van Deurs B (1988) Inhibition of endocytosis from coated pits by acidification of the cytosol. J Cell Biochem 36(1):73–81CrossRefGoogle Scholar
  47. Sehested M, Skovsgaard T, van Deurs B, Winther-Nielsen H (1987a) Increase in nonspecific adsorptive endocytosis in anthracycline- and vinca alkaloid-resistant Ehrlich ascites tumor cell lines. J Natl Cancer Inst 78(1):171–179Google Scholar
  48. Sehested M, Skovsgaard T, van Deurs B, Winther-Nielsen H (1987b) Increased plasma membrane traffic in daunorubicin resistant P388 leukaemic cells. Effect of daunorubicin and verapamil. Br J Cancer 56(6):747–751Google Scholar
  49. Sharom FJ (1997) The P-glycoprotein efflux pump: how does it transport drugs? J Membr Biol 160(3):161–175CrossRefGoogle Scholar
  50. Sirotnak FM, Yang CH, Mines LS, Oribe E, Biedler JL (1986) Markedly altered membrane transport and intracellular binding of vincristine in multidrug-resistant Chinese hamster cells selected for resistance to vinca alkaloids. J Cell Physiol 126(2):266–274CrossRefGoogle Scholar
  51. Spoelstra EC, Westerhoff HV, Dekker H, Lankelma J (1992) Kinetics of daunorubicin transport by P-glycoprotein of intact cancer cells. Eur J Biochem 207(2):567–579CrossRefGoogle Scholar
  52. Stein WD, Cardarelli C, Pastan I, Gottesman MM (1994) Kinetic evidence suggesting that the multidrug transporter differentially handles influx and efflux of its substrates. Mol Pharmacol 45(4):763–772Google Scholar
  53. Ulander J, Haymet AD (2003) Permeation across hydrated DPPC lipid bilayers: simulation of the titrable amphiphilic drug valproic acid. Biophys J 85(6):3475–3484CrossRefGoogle Scholar
  54. Wang S, Wan NC, Harrison J, Miller W, Chuckowree I, Sohal S, Hancox TC, Baker S, Folkes A, Wilson F, Thompson D, Cocks S, Farmer H, Boyce A, Freathy C, Broadbridge J, Scott J, Depledge P, Faint R, Mistry P, Charlton P (2004) Design and synthesis of new templates derived from pyrrolopyrimidine as selective multidrug-resistance-associated protein inhibitors in multidrug resistance. J Med Chem 47(6):1339–1350CrossRefGoogle Scholar
  55. Yamanaka N, Kato T, Nishida K, Fujikawa T, Fukushima M, Ota K (1979) Elevation of serum lipid peroxide level associated with doxorubicin toxicity and its amelioration by [dl]-alpha-tocopheryl acetate or coenzyme Q10 in mouse (doxorubicin, toxicity, lipid peroxide, tocopherol, coenzyme Q10). Cancer Chemother Pharmacol 3(4):223–227CrossRefGoogle Scholar
  56. Zamora JM, Pearce HL, Beck WT (1988) Physical–chemical properties shared by compounds that modulate multidrug resistance in human leukemic cells. Mol Pharmacol 33(4):454–462Google Scholar

Copyright information

© EBSA 2007

Authors and Affiliations

  1. 1.School of Pharmacy and Pharmaceutical Sciences, Drug Delivery GroupUniversity of ManchesterManchesterUK
  2. 2.School of Veterinary Medicine and Science, Sutton Bonington CampusUniversity of NottinghamLeicestershireUK

Personalised recommendations