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Assaying Lysosomal Enzyme Activity in Dictyostelium discoideum

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Dictyostelium discoideum

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2814))

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Abstract

Lysosomes are membrane-enclosed organelles that digest intracellular material. They contain more than 50 different enzymes that can degrade a variety of macromolecules including nucleic acids, proteins, polysaccharides, and lipids. In addition to functioning within lysosomes, lysosomal enzymes are also secreted. Alterations in the levels and activities of lysosomal enzymes dysregulates lysosomes, which can lead to the intralysosomal accumulation of biological material and the development of lysosomal storage diseases (LSDs) in humans. Dictyostelium discoideum has a long history of being used to study the trafficking and functions of lysosomal enzymes. More recently, it has been used as a model system to study several LSDs. In this chapter, we outline the methods for assessing the activity of several lysosomal enzymes in D. discoideum (α-galactosidase, β-galactosidase, α-glucosidase, β-glucosidase, β-N-acetylglucosaminidase, α-mannosidase, cathepsin B, cathepsin D, cathepsin F, palmitoyl protein thioesterase 1, and tripeptidyl peptidase 1).

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Abbreviations

DI:

De-ionized

LIMP-2:

Lysosomal integral membrane protein 2

LSD:

Lysosomal storage disease

M6P:

Mannose-6-phosphate

MES:

2-(N-morpholino)ethanesulfonic acid

SM:

Sussman’s medium

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Author information

Authors and Affiliations

  1. Environmental and Life Sciences Graduate Program, Trent University, Peterborough, ON, Canada

    William D. Kim & Robert J. Huber

  2. Department of Biology, Trent University, Peterborough, ON, Canada

    Adam F. DiGiacinto & Robert J. Huber

Authors
  1. William D. Kim
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  2. Adam F. DiGiacinto
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  3. Robert J. Huber
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Corresponding author

Correspondence to Robert J. Huber .

Editor information

Editors and Affiliations

  1. Laboratory of Cellular and Developmental Biology, NIDDK, NIH, Bethesda, MD, USA

    Alan R. Kimmel

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Kim, W.D., DiGiacinto, A.F., Huber, R.J. (2024). Assaying Lysosomal Enzyme Activity in Dictyostelium discoideum. In: Kimmel, A.R. (eds) Dictyostelium discoideum. Methods in Molecular Biology, vol 2814. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-3894-1_4

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  • DOI: https://doi.org/10.1007/978-1-0716-3894-1_4

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  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-0716-3893-4

  • Online ISBN: 978-1-0716-3894-1

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