Myristyl Trimethyl Ammonium Bromide and Octadecyl Trimethyl Ammonium Bromide Are Surface-Active Small Molecule Dynamin Inhibitors that Block Endocytosis Mediated by Dynamin I or Dynamin II

Quan, Annie, McGeachie, Andrew B., Keating, Damien J., van Dam, Ellen M., Rusak, Jenny, Chau, Ngoc, Malladi, Chandra S., Chen, Chen, McCluskey, Adam, Cousin, Michael A. and Robinson, Phillip J. (2007) Myristyl Trimethyl Ammonium Bromide and Octadecyl Trimethyl Ammonium Bromide Are Surface-Active Small Molecule Dynamin Inhibitors that Block Endocytosis Mediated by Dynamin I or Dynamin II. Molecular Pharmacology, 72 6: 1425-1439. doi:10.1124/mol.107.034207

Author Quan, Annie
McGeachie, Andrew B.
Keating, Damien J.
van Dam, Ellen M.
Rusak, Jenny
Chau, Ngoc
Malladi, Chandra S.
Chen, Chen
McCluskey, Adam
Cousin, Michael A.
Robinson, Phillip J.
Title Myristyl Trimethyl Ammonium Bromide and Octadecyl Trimethyl Ammonium Bromide Are Surface-Active Small Molecule Dynamin Inhibitors that Block Endocytosis Mediated by Dynamin I or Dynamin II
Journal name Molecular Pharmacology   Check publisher's open access policy
ISSN 0026-895X
Publication date 2007-12-01
Year available 2007
Sub-type Article (original research)
DOI 10.1124/mol.107.034207
Volume 72
Issue 6
Start page 1425
End page 1439
Total pages 15
Place of publication Bethesda MD
Publisher American Society for Pharamacology and Experimental Therapeutics
Language eng
Subject 110306 Endocrinology
110201 Cardiology (incl. Cardiovascular Diseases)
111201 Cancer Cell Biology
Abstract Cell Signalling Unit, Children's Medical Research Institute, University of Sydney, Sydney, Australia (A.Q., A.B.M., E.M.v.D., J.R., N.C., C.S.M., P.J.R.); Chemistry, School of Environmental and Life Sciences, University of Newcastle, Newcastle, Australia (A.M.); Department of Human Physiology, Flinders University, Adelaide, Australia (D.J.K.), Prince Henry's Institute of Medical Research, Victoria, Australia (C.C.); and, Membrane Biology Group, Centre for Integrative Physiology, University of Edinburgh, Edinburgh, Scotland (M.A.C) Dynamin is a GTPase enzyme involved in membrane constriction and fission during endocytosis. Phospholipid binding via its pleckstrin homology domain maximally stimulates dynamin activity. We developed a series of surface-active small-molecule inhibitors, such as myristyl trimethyl ammonium bromide (MiTMAB) and octadecyltrimethyl ammonium bromide (OcTMAB), and we now show MiTMAB targets the dynamin-phospholipid interaction. MiTMAB inhibited dynamin GTPase activity, with a Ki of 940 ± 25 nM. It potently inhibited receptor-mediated endocytosis (RME) of transferrin or epidermal growth factor (EGF) in a range of cells without blocking EGF binding, receptor number, or autophosphorylation. RME inhibition was rapidly reversed after washout. The rank order of potency for a variety of MiTMAB analogs on RME matched the rank order for dynamin inhibition, suggesting dynamin recruitment to the membrane is a primary cellular target. MiTMAB also inhibited synaptic vesicle endocytosis in rat brain nerve terminals (synaptosomes) without inducing depolarization or morphological defects. Therefore, the drug rapidly and reversibly blocks multiple forms of endocytosis with no acute cellular damage. The unique mechanism of action of MiTMAB provides an important tool to better understand dynamin-mediated membrane trafficking events in a variety of cells.
References Achiriloaie M, Barylko B, and Albanesi JP (1999) Essential role of the dynamin pleckstrin homology domain in receptor-mediated endocytosis. Mol Cell Biol 19: 1410–1415. Ahyayauch H, Requero MA, Alonso A, Bennouna M, and Goni FM (2002) Surfactant effects of chlorpromazine and imipramine on lipid bilayers containing sphingomyelin and cholesterol. J Colloid Interface Sci 256:284–289. Anggono V, Smillie KJ, Graham ME, Valova VA, Cousin MA, and Robinson PJ (2006) Syndapin I is the phosphorylation-regulated dynamin i partner in synaptic vesicle endocytosis. Nat Neurosci 9:752–760. Artalejo CR, Lemmon MA, Schlessinger J, and Palfrey HC (1997) Specific role for the PH domain of dynamin-1 in the regulation of rapid endocytosis in adrenal chromaffin cells. EMBO J 16:1565–1574. Atkin R, Craig VS, Wanless EJ, and Biggs S (2003) The influence of chain length and electrolyte on the adsorption kinetics of cationic surfactants at the silica-aqueous solution interface. J Colloid Interface Sci 266:236–244. Attwood D, Mosquera V, Garcia M, Suarez MJ, and Sarmiento F (1995) A comparison of the micellar properties of structurally related antidepressant drugs. J Colloid Interface Sci 175:201–262. Atwood WJ (2001) A combination of low-dose chlorpromazine and neutralizing antibodies inhibits the spread of JC virus (JCV) in a tissue culture model: implications for prophylactic and therapeutic treatment of progressive multifocal leukencephalopathy. J Neurovirol 7:307–310. Barylko B, Binns DD, Lin KM, Atkinson ML, Jameson DM, Yin HL, and Albanesi JP (1998) Synergistic activation of dynamin GTPase by Grb2 and phosphoinositides. J Biol Chem 273:3791–3797. Chow RH, von Ruden L, and Neher E (1992) Delay in vesicle fusion revealed by electrochemical monitoring of single secretory events in adrenal chromaffin cells. Nature 356:60–63. Cousin MA and Robinson PJ (1998) Ba2 does not support synaptic vesicle retrieval in rat isolated presynaptic nerve terminals. Neurosci Lett 253:1–4. Cousin MA and Robinson PJ (2000) Ca2 inhibition of dynamin arrests synaptic vesicle recycling at the active zone. J Neurosci 20:949–957. Cousin MA and Robinson PJ (2001) The dephosphins: dephosphorylation by calcineurin triggers synaptic vesicle endocytosis. Trends Neurosci 24:659–665. Damke H, Binns DD, Ueda H, Schmid SL, and Baba T (2001) Dynamin GTPase domain mutants block endocytic vesicle formation at morphologically distinct stages. Mol Biol Cell 12:2578–2589. Davies PJ, Cornwell MM, Johnson JD, Reggianni A, Myers M, and Murtaugh MP (1984) Studies on the effects of dansylcadaverine and related compounds on receptor-mediated endocytosis in cultured cells. Diabetes Care 7 (Suppl 1):35–41. Dunkley PR, Jarvie PE, Heath JW, Kidd GJ, and Rostas JA (1986) A rapid method for isolation of synaptosomes on Percoll gradients. Brain Res 372:115–129. Ferguson SM, Brasnjo G, Hyashi M, Wolfel M, Collesi C, Giovedi S, Raimondi A, Gong LW, Areil P, Paradise S, et al. (2007) A selective activity-dependent requirement for dynamin 1 in synaptic vesicle endocytosis. Science 316:570–574. Fischer H, Gottschlich R, and Seelig A (1998) Blood-brain barrier permeation: molecular parameters governing passive diffusion. J Membr Biol 165:201–211. Ga´borik Z and Hunyady L (2004) Intracellular trafficking of hormone receptors. Trends Endocrinol Metab 15:286–293. Garcia P, Gupta R, Shah S, Morris AJ, Rudge SA, Scarlata S, Petrova V, McLaughlin S, and Rebecchi MJ (1995) The pleckstrin homology domain of phospholipase C-delta1 binds with high affinity to phosphatidylinositol 4,5-bisphosphate in bilayer membranes. Biochemistry 34:16228–16234. Geladopoulos TP, Sotiroudis TG, and Evangelopoulos AE (1991) A malachite green colorimetric assay for protein phosphatase activity. Anal Biochem 192:112–116. Gray JA, Sheffler DJ, Bhatnagar A, Woods JA, Hufeisen SJ, Benovic JL, and Roth BL (2001) Cell-type specific effects of endocytosis inhibitors on 5-hydroxytryptamine2a receptor desensitization and resensitization reveal an arrestin-, GRK2-, and GRK5-independent mode of regulation in human embryonic kidney 293 Cells. Mol Pharmacol 60:1020–1030. Hill TA, Odell LR, Edwards JK, Graham ME, McGeachie AB, Rusak J, Quan A, Abagyan R, Scott JL, Robinson PJ, et al. (2005) Small molecule inhibitors of dynamin i GTPase activity: development of dimeric tyrphostins. J Med Chem 48:7781–7788. Hill TA, Odell LR, Quan A, Abagyan R, Ferguson G, Robinson PJ, and McCluskey A (2004) Long chain amines and long chain ammonium salts as novel inhibitors of dynamin GTPase activity. Bioorg Med Chem Lett 14:3275–3278. Hilliard LM, Osicka TM, Robinson PJ, Nikolic-Paterson DJ, and Comper WD (2006) Characterisation of the urinary degradation pathway in the isolated perfused rat kidney. J Lab Clin Med 147:36–44. Hinshaw JE (2000) Dynamin and its role in membrane fission. Annu Rev Cell Dev Biol 16:483–519. Hohenwallner W and Wimmer E (1973) The malachite green micromethod for the determination of inorganic phosphate. Clin Chim Acta 45:169–175. Jockusch WJ, Praefcke GJ, McMahon HT, and Lagnado L (2005) Clathrin-dependent and clathrin-independent retrieval of synaptic vesicles in retinal bipolar cells. Neuron 46:869–878. Kaksonen M, Toret CP, and Drubin DG (2006) Harnessing actin dynamics for clathrin-mediated endocytosis. Nat Rev Mol Cell Biol 7:404–414. Keating DJ, Rychkov GY, and Roberts ML (2001) Oxygen sensitivity in the sheep adrenal medulla: role of SK channels. Am J Physiol Cell Physiol 281:C1434– C1441. Klein DE, Lee A, Frank DW, Marks MS, and Lemmon MA (1998) The pleckstrin homology domains of dynamin isoforms require oligomerization for high affinity phosphoinositide binding. J Biol Chem 273:27725–27733. Koenig JH and Ikeda K (1989) Disappearance and reformation of synaptic vesicle membrane upon transmitter release observed under reversible blockage of membrane retrieval. J Neurosci 9:3844–3860. Larkin JM, Brown MS, Goldstein JL, and Anderson RG (1983) Depletion of intracellular potassium arrests coated pit formation and receptor-mediated endocytosis in fibroblasts. Cell 33:273–285. Le Roy C and Wrana JL (2005) Clathrin- and non-clathrin-mediated endocytic regulation of cell signalling. Nat Rev Mol Cell Biol 6:112–126. Lee A, Frank DW, Marks MS, and Lemmon MA (1999) Dominant-negative inhibition of receptor-mediated endocytosis by a dynamin-1 mutant with a defective pleckstrin homology domain. Curr Biol 9:261–264. Lemmon MA (2003) Phosphoinositide recognition domains. Traffic 4:201–213. Lin HC, Barylko B, Achiriloaie M, and Albanesi JP (1997) Phosphatidylinositol (4,5)-bisphosphate-dependent activation of dynamins I and II lacking the proline/ arginine-rich domains. J Biol Chem 272:25999–26004. Lindgren CA, Emery DG, and Haydon PG (1997) Intracellular acidification reversibly reduces endocytosis at the neuromuscular junction. J Neurosci 17:3074–3084. Liu JP and Robinson PJ (1995) Dynamin and endocytosis. Endocr Rev 16:590–607. Marks B and McMahon HT (1998) Calcium triggers calcineurin-dependent synaptic vesicle recycling in mammalian nerve terminals. Curr Biol 8:740–749. Marks B, Stowell MH, Vallis Y, Mills IG, Gibson A, Hopkins CR, and McMahon HT 1438 Quan et al. Downloaded from at UQ Library on January 12, 2009 (2001) GTPase activity of dynamin and resulting conformation change are essential for endocytosis. Nature 410:231–235. McNiven MA (1998) Dynamin: a molecular motor with pinchase action. Cell 94:151– 154. Nicholls DG and Sihra TS (1986) Synaptosomes possess an exocytotic pool of glutamate. Nature 321:772–773. Quan A and Robinson PJ (2005) Rapid purification of native dynamin I and colorimetric GTPase assay. Methods Enzymol 404:556–569. Robinson PJ, Sontag J-M, Liu JP, Fykse EM, Slaughter C, McMahon HT, and Su¨ dhof TC (1993) Dynamin GTPase regulated by protein kinase C phosphorylation in nerve terminals. Nature 365:163–166. Roux A, Uyhazi K, Frost A, and De Camilli P (2006) GTP-dependent twisting of dynamin implicates constriction and tension in membrane fission. Nature 441: 528–531. Salim K, Bottomley MJ, Querfurth E, Zvelebil MJ, Gout I, Scaife R, Margolis RL, Gigg R, Smith CI E, Driscoll PC, et al. (1996) Distinct specificity in the recognition of phosphoinositides by the pleckstrin homology domains of dynamin and Bruton’s tyrosine kinase. EMBO J 15:6241–6250. Scaife R, Venien-Bryan C, and Margolis RL (1998) Dual function C-terminal domain of dynamin-1: modulation of self-assembly by interaction of the assembly site with SH3 domains. Biochemistry 37:17673–17679. Schreier S, Malheiros SV and de Paula E (2000) Surface active drugs: self-association and interaction with membranes and surfactants. Physicochemical and biological aspects. Biochim Biophys Acta 1508:210–234. Scurlock JE and Curtis BM (1981) Tetraethylammonium derivatives: ultralongacting local anesthetics? Anesthesiology 54:265–269. Sheff D, Pelletier L, O’Connell CB, Warren G, and Mellman I (2002) Transferrin receptor recycling in the absence of perinuclear recycling endosomes. J Cell Biol 156:797–804. Stowell MH, Marks B, Wigge P, and McMahon HT (1999) Nucleotide-dependent conformational changes in dynamin: evidence for a mechanochemical molecular spring. Nat Cell Biol 1:27–32. Tan TC, Valova VA, Malladi CS, Graham ME, Berven LA, Jupp OJ, Hansra G, McClure SJ, Sarcevic B, Boadle RA, et al. (2003) Cdk5 is essential for synaptic vesicle endocytosis. Nat Cell Biol 5:701–710. Trowbridge IS, Collawn JF, and Hopkins CR (1993) Signal-dependent membrane protein trafficking in the endocytic pathway. Annu Rev Cell Biol 9:129–161. Vallis Y, Wigge P, Marks B, Evans PR, and McMahon HT (1999) Importance of the pleckstrin homology domain of dynamin in clathrin-mediated endocytosis. Curr Biol 9:257–260. van der Bliek AM, Redelmeier TE, Damke H, Tisdale EJ, Meyerowitz EM, and Schmid SL (1993) Mutations in human dynamin block an intermediate stage in coated vesicle formation. J Cell Biol 122:553–563. Van Veldhoven PP and Mannaerts GP (1987) Inorganic and organic phosphate measurements in the nanomolar range. Anal Biochem 161:45–48. Wang L-H, Rothberg KG, and Anderson RG (1993) Mis-assembly of clathrin lattices on endosomes reveals a regulatory switch for coated pit formation. J Cell Biol 123:1107–1117. Yan Z, Chi P, Bibb JA, Ryan TA, and Greengard P (2002) Roscovitine: a novel regulator of P/Q-type calcium channels and transmitter release in central neurons. J Physiol 540:761–770. Zheng J, Cahill SM, Lemmons MA, Fushman D, Schlessinger J, Cowburn D, and Lemmon MA (1996) Identification of the binding site for acidic phospholipids on the PH domains of dynamin: implications for stimulation of GTPase activity. J Mol Biol 255:14–21.
Q-Index Code C1

Document type: Journal Article
Sub-type: Article (original research)
Collections: Excellence in Research Australia (ERA) - Collection
School of Biomedical Sciences Publications
Version Filter Type
Citation counts: TR Web of Science Citation Count  Cited 60 times in Thomson Reuters Web of Science Article | Citations
Scopus Citation Count Cited 60 times in Scopus Article | Citations
Google Scholar Search Google Scholar
Created: Wed, 14 Jan 2009, 02:35:03 EST by Judy Dingwall on behalf of School of Biomedical Sciences