Abstract
Neuronal ceroid lipofuscinoses (NCLs) are the most common hereditary neurodegenerative diseases of childhood. The infantile form, INCL, is caused by lysosomal palmitoyl-protein thioesterase (PPT) deficiency, which impairs the cleavage of thioester linkages in palmitoylated proteins, preventing their hydrolysis by lysosomal proteinases. Consequent accumulation of these lipid-modified proteins (constituents of ceroid) in lysosomes leads to INCL. Because thioester linkages are susceptible to nucleophilic attack, drugs with this property may have therapeutic potential for INCL. We report here that two such drugs, phosphocysteamine and N-acetylcysteine, disrupt thioester linkages in a model thioester compound, [14C]palmitoyl∼CoA. Most importantly, in lymphoblasts derived from INCL patients, phosphocysteamine, a known lysosomotrophic drug, mediates the depletion of lysosomal ceroids, prevents their re-accumulation and inhibits apoptosis. Our results define a novel pharmacological approach to lysosomal ceroid depletion and raise the possibility that nucleophilic drugs such as phosphocysteamine hold therapeutic potential for INCL.
This is a preview of subscription content, access via your institution
Access options
Subscribe to this journal
Receive 12 print issues and online access
$209.00 per year
only $17.42 per issue
Rent or buy this article
Prices vary by article type
from$1.95
to$39.95
Prices may be subject to local taxes which are calculated during checkout
Similar content being viewed by others
References
Schmidt, M.F.G. Fatty acylation of proteins. Biochim. Biophys. Acta 988, 411–426 (1989).
Yalovky, S., Rodriguez-Concepcion, M. & Gruissem, W. Lipid modifications of proteins-slipping in and out of membranes. Trends Plant Res. 4, 439–445 (1999).
Bizzozero, O.A. The mechanism and functional roles of protein palmitoylation in the nervous system. Neuropediatrics 28, 23–26 (1997).
Randall, W.R. Cellular expression of a cloned, hydrophilic, murine acetylcholinesterase. Evidence of palmitoylated membrane bound forms. J. Biol. Chem. 269, 12367–12374 (1994).
Robinson, L.J., Busconi, L. & Michel, T. Agonist-modulated palmitoylation of endothelial nitric oxide synthase. J. Biol. Chem. 270, 995–998 (1995).
Stults, J.T. et al. Lung surfactant protein SP-C from human, bovine, and canine sources contains palmityl cysteine thioester linkages. Am. J. Physiol. 261, L118–L125 (1991).
Hoeg, J.M., Meng, M.S., Ronan, R., Fairwell, T. & Brewer, H.B. Jr. Human apolipoprotein A-I. Post-translational modification by fatty acid acylation. J. Biol. Chem. 261, 3911–3914 (1986).
Huang, G., Lee, D.M. & Singh, S. Identification of the thiol ester linked lipids in apolipoprotein B. Biochemistry 27, 1395–1400 (1988).
Jing, S.Q. & Trowbridge, I.S. Identification of the intermolecular disulfide bonds of the human transferrin receptor and its lipid-attachment site. EMBO J. 6, 327–331 (1987).
Wedegaertner, P.B. & Bourne, H.R. Activation and depalmitoylation of Gsα . Cell 77, 1063–1070 (1994).
Wedegaertner, P.B., Wilson, P.T. & Bourne, H.R. Lipid modification of trimeric G proteins. J. Biol. Chem. 270, 503–506 (1995).
Hancock, J.F., Magee, A.I., Childs, J.E. & Marshall, C.J. All ras proteins are polyisoprenylated but only some are palmitoylated. Cell 57, 1167–1177 (1989).
Camp, L.A. & Hofmann, S.L. Purification and properties of a palmitoyl-protein thioesterase that cleaves palmitate from H-ras. J. Biol. Chem. 268, 22566–22574 (1993).
Camp, L.A., Verkruyse, L.A. Afendis, S.J., Slaughter, C.A. & Hofmann, S.L. Molecular cloning and expression of palmitoyl-protein thioesterase. J. Biol. Chem. 269, 23212–23219 (1994).
Verkruyse, L.A. & Hofmann, S.L. Lysosomal targeting of palmitoyl protein thioesterase. J. Biol. Chem. 271, 15831–15836 (1996).
Hellsten, E., Vesa, J., Olkkonen, V.M., Jalanko, A. & Peltonen, L. Human palmitoyl protein thioesterase: evidence for lysosomal targeting of the enzyme and disturbed cellular routing in infantile neuronal ceroid lipofuscinosis. EMBO J. 15, 5240–5245 (1996).
Sleat, D.E., Sohar, I., Lackland, H., Majercak, J. & Lobel, P. Rat brain contains high levels of mannose-6-phosphorylated glycoproteins including lysosomal enzymes and palmitoyl-protein thioesterase, an enzyme implicated in infantile neuronal lipofuscinosis. J. Biol. Chem. 271, 19191–19198 (1996).
Jarvela, I. Infantile form of neuronal ceroid lipofuscinosis (CLN1) maps to the short arm of chromosome 1. Genomics 9, 170–173 (1991).
Vesa, J. et al. Mutations in palmitoyl-protein thioesterase gene causing infantile neuronal ceroid lipofuscinosis. Nature 376, 584–587 (1995).
Schlesinger, M.J., Veit, M. & Schmidt, M.F.G. in Palmitoylation of cellular and viral proteins. (ed. Schlesinger, M.J.) 1–19 (CRC, Boca Ratan, Florida, 1972).
Lu, J.-Y, Verkruyse, L.A. & Hofmann, S.L. Lipid thioesters derived from acylated proteins accumulate in infantile neuronal ceroid lipofuscinosis: Correction of the defect in lymphoblasts by recombinant palmitoyl-protein thioesterase. Proc. Natl. Acad. Sci. USA 93, 10046–10050 (1996).
Hofmann, S.L., Lee, L.A., Lu, J. -Y. & Verkruyse, L.A. Palmitoyl-protein thioesterase and the molecular pathogenesis of infantile neuronal ceroid lipofuscinosis. Neuropediatrics 28, 27–30 (1997).
Zhang, Z. et al. Palmitoyl-protein thioesterase gene expression in the developing mouse brain and retina: Implications for early loss of vision in infantile neuronal ceroid lipofuscinosis. Gene 231, 203–211 (1999).
Isosomppi, J. et al. Developmental expression of palmitoyl protein thioesterase in normal mice. Brain Res. Dev. Brain Res. 118, 1–11 (1999).
Suopanki, J., Tyynela, J. Baumann, M. & Haltia, M. Palmitoyl-protein thioesterase, an enzyme implicated in neurodegeneration, is localized in neurons and is developmentally regulated in rat brain. Neurosci. Lett. 265, 53–56 (1999).
Jocelyn, P.C. in Biochemistry of the SH groups. 63–93 (Academic Press, New York, 1972).
Bolanowski, M.A., Earles, B.J. & Lennarz, W.J. Fatty acylation of proteins during development of sea urchin embryos. J. Biol. Chem. 259, 4934–4940 (1984).
Broyer, M. et al. Clinical polymorphism of cystinosis encephalopathy. Results of treatment with cysteamine. J. Inherit. Metab. Dis. 19, 65–75 (1996).
Assadi, F.K., Mullin, J.J. & Beckman, D.A. Evaluation of the reproductive and developmental safety of cysteamine in the rat: effects on female reproduction and early embryonic development. Teratology 58, 88–95 (1998).
Das, A.K. et al. Molecular genetics of palmitoyl-protein thioesterase deficiency in the US. J. Clin. Invest. 102, 361–370 (1998).
Waliany, S., Das, A.K., Gaben, A., Wisniewski, K.E. & Hofmann, S.L. Identification of three novel mutations of the palmitoyl-protein thioesterase-1 (PPT1) gene in children with neuronal ceroid lipofucscinosis. Hum. Mutat. 15, 206–207 (2000).
Tyynela, J., Palmer, D.N., Baumann, M. & Haltia, M. Storage of saposins A and D in infantile neuronal ceroid-lipofuscinosis. FEBS Lett . 330, 8–12 (1993).
Kishimoto, Y., Hiraiwa, M. & O'Brien, J.S. Saposins: Structure, function, distribution, and molecular genetics. J. Lipid Res. 33, 1255–1267 (1992).
Cho, S. & Dawson, G. Palmitoyl protein thioesterase 1 protects against apoptosis mediated by Ras-Akt-caspase pathway in neuroblastoma cells. J. Neurochem 74, 1478–1488 (2000).
Cho, S., Dawson, P.E. & Dawson, G. Antisense palmitoyl protein thioesterase 1 (PPT1) treatment inhibits PPT1 activity and increases cell death in LA-N-5 neuroblastoma cells. J. Neurosci. Res. 62, 234–240 (2000).
Riikonen, R., Vanhanen, S.L., Tyynela, J., Santavuori, P. & Turpeinen, U. CSF insulin-like growth factor-1 in infantile neuronal ceroid lipofuscinosis. Neurology 54, 1828–1832 (2000).
Ben-Sasson, S.A., Sherman, Y. & Gavrieli, Y. Identification of dying cells—In Situ Staining. in Methods in Cell Biology Vol. XLVI. (eds. Schwartz, L.M. & Osborne, B.A.) 29–35 (Academic Press, New York, 1995).
Armstrong, D., Koppang, N. & Rider, J.A. Ceroid Lipofuscinosis (Batten's Disease) (Elsevier, Amsterdam, The Netherlands, 1982).
Goebel, H.H., Mole, S.E. & Lake, B.D., eds. The neuronal ceroid lipofuscinoses (Batten's disease). in Biomedical and Health Res. 211 (IOS Press, Amsterdam, The Netherlands, 1999).
Rapola, J. & Haltia, M. Cytoplasmic inclusions in the vermiform appendix and skeletal muscle in two types of so-called neuronal ceroid-lipofuscinosis. Brain 96, 833–840 (1973).
Santavuori, P. Neuronal ceroid-lipofuscinosis in childhood. Brain Dev. 10, 80–83 (1988).
Wisniewski, K.E., Kida, E., Patxot, O.F. & Connell, F. Variability in the clinical and pathological findings in the neuronal ceroid lipofuscinoses: Review of data and observations. Am. J. Med. Genet. 42, 525–532 (1992).
Wisniewski, K.E. et al. Palmitoyl-protein thioesterase deficiency in a novel granular variant of LINCL. Pediatr. Neurol. 18, 119–123 (1998).
Bellizzi, J.J. 3rd et al. The crystal structure of palmitoyl protein thioesterase 1 and the molecular basis of infantile neuronal ceroid lipofuscinosis. Proc. Natl. Acad. Sci. USA 97, 4573–4578 (2000).
Gahl, W.A., Schneider, J.A. & Aula, P.P. Cystinosis: A disorder of lysosomal membrane transport. in The Metabolic and Molecular Bases of Inherited Disease, Vol. III, 7th ed. (eds. Scriver, C.R., Beaudet, A.L., Sly, W.S. & Valle, D.) 3763–3782 (McGraw-Hill, New York, 1995).
Zhang, Z. et al. Severe fibronectin-deposit renal glomerular disease in mice lacking uteroglobin. Science 276, 1408–1412 (1997).
Zheng, F., Kundu, G.C., Zhang, Z., Ward, J., DeMayo, F. and Mukherjee, A.B. Uteroglobin is essential in preventing immunoglobulin A nephropathy in mice. Nature Med. 5, 1018–1025 (1999).
Acknowledgements
We thank I. Owens, J.Y. Chou and J.B. Sidbury Jr for critical review of the manuscript; W. Gahl for discussions during this study; M. Hiraiwa for a gift of saposin antibodies; and S. Everett and R. Dreyfuss for technical assistance with photomicrography. This study was supported in part by a `Bench-to-Bedside Award 1999' (to A.B.M) from the Clinical Center of the NIH and by a grant #RO1 NS 389 88-01 (to K.E.W) from the National Institute of Neurological Disorders and Stroke.
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Zhang, Z., Butler, J., Levin, S. et al. Lysosomal ceroid depletion by drugs: Therapeutic implications for a hereditary neurodegenerative disease of childhood. Nat Med 7, 478–484 (2001). https://doi.org/10.1038/86554
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1038/86554
This article is cited by
-
Neuronal Ceroid Lipofuscinosis: Potential for Targeted Therapy
Drugs (2021)
-
Emerging new roles of the lysosome and neuronal ceroid lipofuscinoses
Molecular Neurodegeneration (2019)
-
Current and Emerging Treatment Strategies for Neuronal Ceroid Lipofuscinoses
CNS Drugs (2019)
-
Noninvasive targeting delivery and in vivo magnetic resonance tracking method for live apoptotic cells in cerebral ischemia with functional Fe2O3 magnetic nanoparticles
Journal of Nanobiotechnology (2016)
-
Moving towards effective therapeutic strategies for Neuronal Ceroid Lipofuscinosis
Orphanet Journal of Rare Diseases (2016)