Download PDF
Thromb Haemost 2017; 117(01): 105-115
DOI: 10.1160/TH16-01-0045
Cellular Haemostasis and Platelets
Schattauer GmbH

Several adaptor proteins promote intracellular localisation of the transporter MRP4/ABCC4 in platelets and haematopoietic cells

Yvonne Schaletzki
1   Department of Pharmacology, Center of Drug Absorption and Transport (C_DAT), University Medicine Greifswald, Greifswald, Germany
,
Marie-Luise Kromrey
1   Department of Pharmacology, Center of Drug Absorption and Transport (C_DAT), University Medicine Greifswald, Greifswald, Germany
,
Susanne Bröderdorf
1   Department of Pharmacology, Center of Drug Absorption and Transport (C_DAT), University Medicine Greifswald, Greifswald, Germany
,
Elke Hammer
2   Department of Functional Genomics, Interfaculty Institute of Genetics and Functional Genomics, University Medicine Greifswald, Greifswald, Germany
,
Markus Grube
1   Department of Pharmacology, Center of Drug Absorption and Transport (C_DAT), University Medicine Greifswald, Greifswald, Germany
,
Paul Hagen
1   Department of Pharmacology, Center of Drug Absorption and Transport (C_DAT), University Medicine Greifswald, Greifswald, Germany
,
Sonja Sucic
3   Department of Pharmacology, Center of Physiology and Pharmacology, Medical University of Vienna, Vienna, Austria
,
Michael Freissmuth
3   Department of Pharmacology, Center of Physiology and Pharmacology, Medical University of Vienna, Vienna, Austria
,
Uwe Völker
2   Department of Functional Genomics, Interfaculty Institute of Genetics and Functional Genomics, University Medicine Greifswald, Greifswald, Germany
,
Andreas Greinacher
4   Department of Immunology and Transfusion Medicine, University Medicine Greifswald, Greifswald, Germany
,
Bernhard H. Rauch
1   Department of Pharmacology, Center of Drug Absorption and Transport (C_DAT), University Medicine Greifswald, Greifswald, Germany
,
Heyo K. Kroemer   *
1   Department of Pharmacology, Center of Drug Absorption and Transport (C_DAT), University Medicine Greifswald, Greifswald, Germany
,
Gabriele Jedlitschky
1   Department of Pharmacology, Center of Drug Absorption and Transport (C_DAT), University Medicine Greifswald, Greifswald, Germany
› Author Affiliations Financial support: The work was supported by the Deutsche Forschungsgemeinschaft (DFG) through grant JE 234/4–1 to GJ and by the European Union through grant EU-FP7-REGPOT-2010–1 (“EnVision”, grant agreement no. 264143).
Further Information

Publication History

Received: 18 January 2016

Accepted after major revision: 20 September 2016

Publication Date:
01 December 2017 (online)

Also available at
    Permissions and Reprints

Summary

The multidrug resistance protein 4 (MRP4/ABCC4) has been identified as an important transporter for signalling molecules including cyclic nucleotides and several lipid mediators in platelets and may thus represent a novel target to interfere with platelet function. Besides its localisation in the plasma membrane, MRP4 has been also detected in the membrane of dense granules in resting platelets. In polarised cells it is localised at the basolateral or apical plasma membrane. To date, the mechanism of MRP4 trafficking has not been elucidated; protein interactions may regulate both the localisation and function of this transporter. We approached this issue by searching for interacting proteins by in vitro binding assays, followed by immunoblotting and mass spectrometry, and by visualising their co-localisation in platelets and haematopoietic cells. We identified the PDZ domain containing scaffold proteins ezrin-binding protein 50 (EBP50/NHERF1), postsynaptic density protein 95 (PSD95), and sorting nexin 27 (SNX27), but also the adaptor protein complex 3 subunit β3A (AP3B1) and the heat shock protein HSP90 as putative interaction partners of MRP4. The knockdown of SNX27, PSD95, and AP3B1 by siRNA in megakaryoblastic leuk aemia cells led to a redistribution of MRP4 from intracellular structures to the plasma membrane. Inhibition of HSP90 led to a diminished expression and retention of MRP4 in the endoplasmic reticulum. These results indicate that MRP4 localisation and function are regulated by multiple protein interactions. Changes in the adaptor proteins can hence lead to altered localisation and function of the transporter.

Supplementary Material to this article is available at www.thrombosis-online.com.

Keywords

Protein trafficking - PDZ domain - AP3B1 - HSP90 - delta-storage pool deficiencies

* Present address: University Medicine Göttingen, Göttingen, Germany.


 

  • References

  • 1 Ritter CA, Jedlitschky G, Meyer zu Schwabedissen H. et al. Cellular export of drugs and signalling molecules by the ATP-binding cassette transporters MRP4 (ABCC4) and MRP5 (ABCC5). Drug Metab Rev 2005; 37: 253-278.
    CrossrefPubMedSearch in Google Scholar
  • 2 Jedlitschky G, Greinacher A, Kroemer HK. Transporters in human platelets: physiologic function and impact for pharmacotherapy. Blood 2012; 119: 3394-3402.
    CrossrefPubMedSearch in Google Scholar
  • 3 Sassi Y, Lipskaia L, Vandecasteele G. et al. Multidrug resistance-associated protein 4 regulates cAMP-dependent signalling pathways and controls human and rat SMC proliferation. J Clin Invest 2008; 118: 2747-2757.
    CrossrefPubMedSearch in Google Scholar
  • 4 Coepsel S, Garcia C, Diez F. et al. Multidrug resistance protein 4 (MRP4/ABCC4) regulates cAMP cellular levels and controls human leukemia cell proliferation and differentiation. J Biol Chem 2011; 286: 6979-6988.
    CrossrefPubMedSearch in Google Scholar
  • 5 Jedlitschky G, Tirschmann K, Lubenow LE. et al. The nucleotide transporter MRP4 (ABCC4) is highly expressed in human platelets and present in dense granules, indicating a role in mediator storage. Blood 2004; 104: 3603-3610.
    CrossrefPubMedSearch in Google Scholar
  • 6 Rius M, Hummel-Eisenbeiss J, Keppler DJ. ATP-dependent transport of leukotrienes B4 and C4 by the multidrug resistance protein ABCC4 (MRP4). Pharmacol Exp Ther 2008; 324: 86-94.
    PubMedSearch in Google Scholar
  • 7 Jedlitschky G, Cattaneo M, Lubenow LE. et al. Role of MRP4 (ABCC4) in platelet adenine nucleotide-storage: evidence from patients with delta-storage pool deficiencies. Am J Pathol 2010; 176: 1097-1103.
    CrossrefPubMedSearch in Google Scholar
  • 8 Niessen J, Jedlitschky G, Grube M. et al. Expression of ABC-type transport proteins in human platelets. Pharmacogenet Genomics 2010; 20: 396-400.
    CrossrefPubMedSearch in Google Scholar
  • 9 Mattiello T, Guerriero R, Lotti LV. et al. Aspirin extrusion from human platelets through multidrug resistance protein-4-mediated transport evidence of a reduced drug action in patients after coronary artery bypass grafting. J Am Coll Cardiol 2011; 58: 752-761.
    CrossrefPubMedSearch in Google Scholar
  • 10 Borgognone A, Pulcinelli FM. Reduction of cAMP and cGMP inhibitory effects in human platelets by MRP4-mediated transport. Thromb Haemost 2012; 108: 955-962.
    Thieme ConnectPubMedSearch in Google Scholar
  • 11 Ulrych T, Böhm A, Polzin A. et al. Release of sphingosine-1-phosphate from human platelets is dependent on thromboxane formation. J Thromb Haemost 2011; 09: 790-798.
    CrossrefPubMedSearch in Google Scholar
  • 12 Decouture B, Dreano E, Belleville-Rolland T. et al. Impaired platelet activation and cAMP homeostasis in MRP4-deficient mice. Blood 2015; 126: 1823-1830.
    CrossrefPubMedSearch in Google Scholar
  • 13 Cheepala SB, Pitre A, Fukuda Y. et al. The ABCC4 membrane transporter modulates platelet aggregation. Blood 2015; 126: 2307-2319.
    CrossrefPubMedSearch in Google Scholar
  • 14 Lien LM, Chen ZC, Chung CL. et al. Multidrug resistance protein 4 (MRP4/ABCC4) regulates thrombus formation in vitro and in vivo. Eur J Pharmacol 2014; 737: 159-167.
    CrossrefPubMedSearch in Google Scholar
  • 15 Cattaneo M. Inherited, platelet-based bleeding disorders. J Thromb Haemost 2003; 12: 1628-1636.
    PubMedSearch in Google Scholar
  • 16 Hermansky F, Pudlak P. Albinism associated with hemorrhagic diathesis and unusual pigmented reticular cells in the bone marrow. Blood 1959; 14: 162-169.
    PubMedSearch in Google Scholar
  • 17 Hoque MT, Cole SP. Down-regulation of Na+/H+ exchanger regulatory factor 1 increases expression and function of multidrug resistance protein 4. Cancer Res 2008; 68: 4802-4809.
    CrossrefPubMedSearch in Google Scholar
  • 18 Park J, Kwak JO, Riederer B. et al. Na+/H+ exchanger regulatory factor 3 is critical for multidrug resistance protein 4-mediated drug efflux in the kidney. J Am Soc Nephrol 2014; 25: 726-736.
    CrossrefPubMedSearch in Google Scholar
  • 19 Hayashi H, Naoi S, Nakagawa T. et al. Sorting nexin 27 interacts with multidrug resistance-associated protein 4 (MRP4) and mediates internalisation of MRP4. J Biol Chem 2012; 287: 15054-15065.
    CrossrefPubMedSearch in Google Scholar
  • 20 Dell’Angelica EC. AP-3-dependent trafficking and disease: the first decade. Curr Opin Cell Biol 2009; 21: 552-559.
    CrossrefPubMedSearch in Google Scholar
  • 21 Jung J, Bohn G, Allroth A. et al. Identification of a homozygous deletion in the AP3B1 gene causing Hermansky-Pudlak syndrome, type 2. Blood 2006; 108: 362-369.
    CrossrefPubMedSearch in Google Scholar
  • 22 Oevermann L, Scheitz J, Starke K. et al. Haematopoietic stem cell differentiation affects expression and function of MRP4 (ABCC4), a transport protein for signalling molecules and drugs. Int J Cancer 2009; 124: 2303-2311.
    CrossrefPubMedSearch in Google Scholar
  • 23 Hermann A, Rauch BH, Braun M. et al. Platelet CD40 ligand (CD40L) - subcellular localisation, regulation of expression, and inhibition by clopidogrel. Platelets 2001; 12: 74-82.
    CrossrefPubMedSearch in Google Scholar
  • 24 Stannek L, Thiele MJ, Ischebeck T. et al. Evidence for synergistic control of glutamate biosynthesis by glutamate dehydrogenases and glutamate in Bacillus subtilis. Environ Microbiol 2015; 17: 3379-3390.
    CrossrefPubMedSearch in Google Scholar
  • 25 Rius M, Nies AT, Hummel-Eisenbeiss J. et al. Cotransport of reduced glutathione with bile salts by MRP4 (ABCC4) localized to the basolateral hepatocyte membrane. Hepatology 2003; 38: 374-384.
    PubMedSearch in Google Scholar
  • 26 Köck K, Koenen A, Giese B. et al. Rapid modulation of the organic anion transporting polypeptide 2B1 (OATP2B1, SLCO2B1) function by protein kinase C-mediated internalisation. J Biol Chem 2010; 285: 11336-11347.
    CrossrefPubMedSearch in Google Scholar
  • 27 Taldone Tl, Sun W, Chiosis G. Discovery and development of heat shock protein 90 inhibitors. Bioorg Med Chem 2009; 17: 2225-2235.
    CrossrefPubMedSearch in Google Scholar
  • 28 Sugiura Tl, Shimizu T, Kijima A. et al. PDZ adaptors: their regulation of epithelial transporters and involvement in human diseases. J Pharm Sci 2011; 100: 3620-3635.
    CrossrefPubMedSearch in Google Scholar
  • 29 Lee K, Klein-Szanto AJ, Kruh GD. Analysis of the MRP4 drug resistance profile in transfected NIH3T3 cells. J Natl Cancer Inst 2000; 92: 1934-1940.
    CrossrefPubMedSearch in Google Scholar
  • 30 van Aubel RA, Smeets PH, Peters JG. et al. The MRP4/ABCC4 gene encodes a novel apical organic anion transporter in human kidney proximal tubules: putative efflux pump for urinary cAMP and cGMP. J Am Soc Nephrol 2002; 13: 595-603.
    PubMedSearch in Google Scholar
  • 31 Niessen J, Jedlitschky G, Grube M. et al. Subfractionation and purification of intracellular granule-structures of human platelets: an improved method based on magnetic sorting. J Immunol Methods 2007; 328: 89-96.
    CrossrefPubMedSearch in Google Scholar
  • 32 Short DB, Trotter KW, Reczek DJ. et al. An apical pdz protein anchors the cystic fibrosis transmembrane conductance regulator to the cytoskeleton. J Biol Chem 1998; 273: 19797-19801.
    CrossrefPubMedSearch in Google Scholar
  • 33 Swiatecka-Urban A, Duhaime M, Coutermarsh B. et al. Pdz domain interaction controls the endocytic recycling of the cystic fibrosis transmembrane conductance regulator. J Biol Chem 2002; 277: 40099-40105.
    CrossrefPubMedSearch in Google Scholar
  • 34 Fanning AS, Anderson JM. Protein modules as organizers of membrane structure. Curr Opin Cell Biol 1999; 11: 432-439.
    CrossrefPubMedSearch in Google Scholar
  • 35 Li C, Krishnamurthy PC, Penmatsa H. et al. Spatiotemporal coupling of cAMP transporter to CFTR chloride channel function in the gut epithelia. Cell 2007; 131: 940-951.
    CrossrefPubMedSearch in Google Scholar
  • 36 Moon C, Zhang W, Ren A. et al. Compartmentalized accumulation of cAMP near complexes of multidrug resistance protein 4 (MRP4) and cystic fibrosis transmembrane conductance regulator (CFTR) contributes to drug-induced diarrhea. J Biol Chem 2015; 290: 11246-11257.
    CrossrefPubMedSearch in Google Scholar
  • 37 de Bartolomeis A, Tomasetti C. Calcium-dependent networks in dopamine-glutamate interaction: the role of postsynaptic scaffolding proteins. Mol Neurobiol 2012; 46: 275-296.
    CrossrefPubMedSearch in Google Scholar
  • 38 Reed GL, Fitzgerald ML, Polgar J. Molecular mechanisms of platelet exocytosis: insights into the “secrete” life of thrombocytes. Blood 2000; 96: 3334-3342.
    PubMedSearch in Google Scholar
  • 39 Ambrosio AL, Boyle JA, Di Pietro SM. Mechanism of platelet dense granule biogenesis: study of cargo transport and function of Rab32 and Rab38 in a model system. Blood 2012; 120: 4072-4081.
    CrossrefPubMedSearch in Google Scholar
  • 40 Khurana N, Bhattacharyya S. Hsp90, the concertmaster: tuning transcription. Front Oncol 2015; 05: 100
    PubMedSearch in Google Scholar
  • 41 Suttitanamongkol S, Gear AR, Polanowska-Grabowska R. Geldanamycin disrupts platelet-membrane structure, leading to membrane permeabilisation and inhibition of platelet aggregation. Biochem J 2000; 345: 307-314.
    CrossrefPubMedSearch in Google Scholar
  • 42 Lalo U, Jones S, Roberts JA, Mahaut-Smith MP, Evans RJ. Heat shock protein 90 inhibitors reduce trafficking of ATP-gated P2×1 receptors and human platelet responsiveness. J Biol Chem 2012; 287: 32747-32754.
    CrossrefPubMedSearch in Google Scholar