Biochemistry and significance of P-type ATPase-dependent lipid transport


Lipid asymmetry is a universal feature of plasma membranes, yet its regulation and significance for the functioning of cells is poorly understood. Hence, identification of the lipid translocases responsible for creating lipid asymmetry is a hot topic in current cell biology. We recently identified two novel P-type ATPases required for inward-directed lipid transport across the plasma membrane in yeast. Moreover, we uncovered a functional link between P-type ATPase-controlled lipid asymmetry and the formation of endocytic vesicles. These findings generated a unique starting point for an interdisciplinary approach to unravel the working mechanisms of lipid translocases and their biological roles.
To this end, the present proposal aims to achieve four key objectives. The first one concerns the fact that P-type ATPases generally function as transporters of cations. Apparently, the ones we identified form an exception and serve as inward-directed lipid pumps. To verify this concept, the newly-identified ATPases will be purified and reconstituted into proteoliposomes and their lipid-transporting activities examined under chemically-defined conditions. Second, proteoliposomes will be fused to giant liposomes and analysed by optical microscopy to explore the role of ATPase-dependent lipid pumping in membrane deformation and vesiculation. Third, our ATPases belong to a subfamily whose members display significant differences in lipid specificity. To map protein regions involved in lipid binding and recognition, hybrid molecules composed of different family members will be generated and their transport functions analysed in vivo. Fourth, since multiple family members localize to Golgi and endosomal compartments, we developed a method to establish whether lipid topology and vesicle budding in these organelles are also subject to P-type ATPase-dependent regulation.
Homologous of the above P-type ATPases have been linked to defective bile secretion and neurological disorders in humans. Apart from providing insight into the fundamental nature of lipid translocation, our investigations should help elucidate the molecular basis of these diseases.



  • J.C.M. Holthuis(2004): The Golgi: a transition point in membrane lipid composition and topology  17 december 2004
  • K. Huitema(2005): The sphingomyelin synthase family  21 november 2005
  • F.G. Tafesse(2009): A Ceramide Sensor Hiding in a Family of Sphingomyelin Synthases  14 december 2009
  • P.M. Verhulst(2009): Flip or Flop: Functional Analysis of a Disease-related Class of Lipid Pumps  14 januari 2009
  • A.M. Vacaru(2009): Mechanisms and Impact of Ceramide Phosphoethanolamine Biosynthesis  14 december 2009

Hoofdstuk in boek

  • J.C.M. Holthuis, J. Jantti, S. Keranen(2004): Topics in Current Genetics. Vol. 10. pp. 39 - 64 , Berlin

Wetenschappelijk artikel

  • Q. Lisman, D. Urli-Stam, T. Pomorski, J.C.M. Holthuis, W. de Cock van Delwijnen, C. Vogelzangs(2004): Protein sorting in the late Golgi of Saccharomyces cerevisiae does not require mannosylated sphingolipids Journal of Biological Chemistry pp. 1020 - 1029
  • J.C.M. Holthuis, J.F. Brouwers, K. Huitema, J. van den Dikkenberg(2004): Identification of a family of animal sphingomyelin synthases EMBO Journal pp. 33 - 44
  • J.C.M. Holthuis, A. Herrmann, G. van Meer, T. Pomorski(2004): Tracking down lipid flippases and their biological functions Journal of Cell Science pp. 805 - 813
  • Q. Lisman, J.C.M. Holthuis, D. Urli-Stam(2004): HOR7, a multi-copy suppressor of the Ca2+-sensitive growth defect in sphingolipid mannosyltransferase-deficient yeast Journal of Biological Chemistry pp. 36390 - 36396
  • J.C.M. Holthuis, G. Lenoir(2004): The elusive flippases Current Biologie pp. R912 - 913
  • T.P. Levine, J.C.M. Holthuis(2005): Lipid traffic: floppy drives and a superhighway Nature Reviews Molecular Cell Biology pp. 209 - 220
  • J.C.M. Holthuis, Q Lisman, T Pomorski, L Luong, N Alder-Baerens(2006): Loss of P4 ATPases Drs2p and Dnf3p disrupts aminophospholipid transport and asymmetry in yeast post-Golgi secretory vesicles Molecular Biology of the Cell pp. 1632 - 1642
  • F. Geta Tafesse, P. Ternes, J.C.M. Holthuis(2006): The multigenic sphingomyelin synthase family Journal of Biological Chemistry pp. 29421 - 29425
  • C. Luberto, D.A. Persaud-Sawin, S.M. Jazwinski, A. Bielawski, T Mousallem, A. Zucker, A. Schulz, R.M. Boustany, L. Kozhaya, G.S. Dbaibo, J.C.M. Holthuis, J. Bielawski, M. Venkaramani(2006): The CLN9 protein, a regulator of dihydroceramide synthase Journal of Biological Chemistry pp. 2784 - 2794
  • A. Uphoff, F. Geta Tafesse, J. van den Dikkenberg, K. Huitema, J.C.M. Holthuis, P. Somerharju, M. Hermansson, S. van der Poel(2007): Both sphingomyelin synthases SMS1 and SMS2 are required for sphingomyelin homeostasis and growth in human HeLa cells Journal of Biological Chemistry pp. 17537 - 17547
  • J.C.M. Holthuis, G. Lenoir, P. Williamson(2007): Origin of lipid asymmetry: the flip side of ion transport. Current Opinion in Chemical Biology pp. 654 - 661
  • B.M. Humbel, C. Tomova, R. Entzeroth, A.J. Verkleij, J.C.M. Holthuis, W.J. Geerts(2009): Membrane contact sites between apicoplast and ER in Toxoplasma gondii revealed by Electron Tomography Traffic pp. 1471 - 1480
  • P. Williamson, G. Lenoir, J.C.M. Holthuis, C.F. Puts(2009): Cdc50p plays a vital role in the ATPase reaction cycle of the putative aminophospholipid transporter Drs2p. Journal of Biological Chemistry pp. 17956 - 17967
  • E. Krieger, L.W. Klomp, L.M. Van der Velden, J.S. Stapelbroek, S.F.J. van de Graaf, R. Berger, P.M. Verhulst, J.C.M. Holthuis, P.V.E. van den Berghe, R.H.J. Houwen(2009): Folding defects in ATP8B1 associated with hereditary cholestasis are ameliorated by 4-phenylbutyrate. Hepatology
  • V. Kondylis, P. Somerharju, J.F. Brouwers, P. Ternes, J.C.M. Holthuis, C. Rabouille, A.M. Vacaru, M. Hermansson, F.G. Tafesse(2009): Sphingomyelin synthase-related protein SMSr controls ceramide homeostasis in the ER Journal of Cell Biology pp. 1013 - 1027
  • J. van den Dikkenberg, P. Ternes, J.C.M. Holthuis, J.F. Brouwers(2009): Sphingomyelin synthase SMS2 displays dual activity as ceramide phosphoethanolamine synthase. Journal of Lipid Research pp. 2270 - 2277
  • P. Williamson, C.F. Puts, J.C.M. Holthuis, J. Krijgsveld, G. Lenoir(2009): In vivo cross-linking of a P4-ATPase protein interaction network reveals a link between aminophospholipid transport and phosphoinositide metabolism Journal of Proteome Research
  • J.C.M. Holthuis, C.F. Puts(2009): Mechanism and significance of P4 ATPase-catalysed lipid transport: lessons from a Na+/K+-pump. BBA Molecular and Cellular Biology of Lipids pp. 603 - 611
  • V Oorschot, RH Houwen, EE van Faassen, JC Holthuis, LW Klomp, TG Pomorski, J Klumperman, LM van der Velden, PM Verhulst(2010): A flippase-independent function of ATP8B1, the protein affected in familial intrahepatic cholestasis type 1, is required for apical protein expression and microvillus formation in polarized epithelial cells Hepatology pp. 2049 - 2060

Professionele publicatie

  • J.C.M. Holthuis, A.I.P.M. De Kroon: Royal Academy Colloquium and Master Class "Lipids moving center stage"


  • B. Qualmann, J.C.M. Holthuis(2005): Mini-symposium "Membrane Traffic", European Life Science Organization Meeting, 3-7 September 2005, Dresden, Germany
  • J.C.M. Holthuis(2006): Invited lectures & organization international scientific meetings





Dr. J.C.M. Holthuis

Verbonden aan

Universität Osnabrück, Fachbereich of Biologie/Chemie


Dr. G. Lenoir


01/01/2004 tot 08/12/2009