Cited by 23Protease and Inhibitor MechanismsProtein Structure and DynamicsLipid Membrane Structure and Behavior

Non-canonical Shedding of TNFα by SPPL2a Is Determined by the Conformational Flexibility of Its Transmembrane Helix

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Spitz, Charlotte, Schlosser, Christine, Guschtschin-Schmidt, Nadja, Stelzer, Walter, Menig, Simon, Götz, Alexander, Haug-Kröper, Martina, Scharnagl, Christina +3 more·2020·iScience

Abstract

Ectodomain (EC) shedding defines the proteolytic removal of a membrane protein EC and acts as an important molecular switch in signaling and other cellular processes. Using tumor necrosis factor (TNF)α as a model substrate, we identify a non-canonical shedding activity of SPPL2a, an intramembrane cleaving aspartyl protease of the GxGD type. Proline insertions in the TNFα transmembrane (TM) helix strongly increased SPPL2a non-canonical shedding, while leucine mutations decreased this cleavage. Using biophysical and structural analysis, as well as molecular dynamic simulations, we identified a flexible region in the center of the TNFα wildtype TM domain, which plays an important role in the processing of TNFα by SPPL2a. This study combines molecular biology, biochemistry, and biophysics to provide insights into the dynamic architecture of a substrate's TM helix and its impact on non-canonical shedding. Thus, these data will provide the basis to identify further physiological substrates of non-canonical shedding in the future.

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Crossref · 10.1016/j.isci.2020.101775

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  • The protein encoded by SPPL2A is involved in noncanonical shedding of TNF-α, and PGRN has been described as a TNF receptor ligand and an antagonist of TNF-α signaling.

    “The protein encoded by SPPL2A is involved in noncanonical shedding of TNF-α34, and PGRN has been described as a TNF receptor ligand and an antagonist of TNF-α signaling35.”

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Authors
Spitz, C., Schlosser, C., Guschtschin-Schmidt, N., Stelzer, W., Menig, S., Götz, A., Haug-Kröper, M., Scharnagl, C., Langosch, D., Muhle-Goll, C., & Fluhrer, R.
Affiliations
  • University of Augsburg (DE)
  • Karlsruhe Institute of Technology (DE)
  • Technical University of Munich (DE)
  • Leibniz Supercomputing Centre (DE)
Corresponding author
Fluhrer, Regina
Venue
iScience
Publisher
Elsevier BV
Published
2020-12
Volume
23
Issue
12
Pages
101775
ISSN
2589-0042 (ISSN-L)
OpenAlex
W3097208119
Cited by
23
References
81
Citations by year
2026: 2 · 2025: 3 · 2024: 5 · 2023: 4 · 2022: 8 · 2021: 1
FWCI
1.11
Type
journal article
Language
en
Open access
Locations
  • iScience (publishedVersion, cc-by-nc-nd)
  • PubMed Central (publishedVersion, cc-by-nc-nd)
  • Repository KITopen (Karlsruhe Institute of Technology) (publishedVersion, cc-by-nc-nd)
  • PubMed (publishedVersion)
  • OPUS (Augsburg University) (submittedVersion, cc-by-nc-nd)
  • DOAJ (DOAJ: Directory of Open Access Journals) (submittedVersion, cc-by-sa)
  • DZNE Pub (submittedVersion, cc-by-nc-nd)
  • KITopen
Funders
  • Deutsche Forschungsgemeinschaft (254872893/FL 635/2-2, 263531414/FOR 2290)
  • Liaoning Medical University
Topics
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Cancer Research › Protease and Inhibitor Mechanisms
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Protein Structure and Dynamics
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Lipid Membrane Structure and Behavior
Keywords
ectodomain shedding, SPPL2a, TNF-α, proline insertion, membrane protein processing, substrate specificity

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