Widespread Expression of a Membrane-Tethered Version of the Soluble Lysosomal Enzyme Palmitoyl Protein Thioesterase-1

  • Charles Shyng
  • Shannon L. Macauley
  • Joshua T. Dearborn
  • Mark S. SandsEmail author
Research Report
Part of the JIMD Reports book series (JIMD, volume 36)


“Cross-correction,” the transfer of soluble lysosomal enzymes between neighboring cells, forms the foundation for therapeutics of lysosomal storage disorders (LSDs). However, “cross-correction” poses a significant barrier to studying the role of specific cell types in LSD pathogenesis. By expressing the native enzyme in only one cell type, neighboring cell types are invariably corrected. In this study, we present a strategy to limit “cross-correction” of palmitoyl-protein thioesterase-1(PPT1), a lysosomal hydrolase deficient in Infantile Neuronal Ceroid Lipofuscinosis (INCL, Infantile Batten disease) to the lysosomal membrane via the C-terminus of lysosomal associated membrane protein-1 (LAMP1). Tethering PPT1 to the lysosomal membrane prevented “cross-correction” while allowing PPT1 to retain its enzymatic function and localization in vitro. A transgenic line harboring the lysosomal membrane-tethered PPT1 was then generated. We show that expression of lysosome-restricted PPT1 in vivo largely rescues the INCL biochemical, histological, and functional phenotype. These data suggest that lysosomal tethering of PPT1 via the C-terminus of LAMP1 is a viable strategy and that this general approach can be used to study the role of specific cell types in INCL pathogenesis, as well as other LSDs. Ultimately, understanding the role of specific cell types in the disease progression of LSDs will help guide the development of more targeted therapeutics.

One Sentence Synopsis: Tethering PPT1 to the lysosomal membrane is a viable strategy to prevent “cross-correction” and will allow for the study of specific cellular contributions in INCL pathogenesis.


Infantile Batten disease Lysosomal storage disorders Neuronal ceroid lipofuscinosis Palmitoyl-protein thioesterase-1 Soluble lysosomal enzymes 



We thank J. Michael White (Transgenic Knockout Micro-Injection Core, WUSTL) for his help with generating the transgenic founders. We thank Dr. Anne Hennig (Vision Research Core, WUSTL) for her help with the electroretinography. In addition, we thank Dr. Bruno Benitez for his advice and technical assistance.


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Copyright information

© SSIEM and Springer-Verlag Berlin Heidelberg 2017

Authors and Affiliations

  • Charles Shyng
    • 1
  • Shannon L. Macauley
    • 2
  • Joshua T. Dearborn
    • 1
  • Mark S. Sands
    • 1
    • 3
    • 4
    Email author
  1. 1.Department of Internal MedicineWashington University School of MedicineSt. LouisUSA
  2. 2.Department of NeurologyWashington University School of MedicineSt. LouisUSA
  3. 3.Department of GeneticsWashington University School of MedicineSt. LouisUSA
  4. 4.Hope Center for Neurological DisordersWashington University School of MedicineSt. LouisUSA

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