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

Spitz, Charlotte; Schlosser, Christine; Guschtschin-Schmidt, Nadja ORCID iD icon 1,2; Stelzer, Walter; Menig, Simon; Götz, Alexander; Haug-Kröper, Martina; Scharnagl, Christina; Langosch, Dieter; Muhle-Goll, Claudia ORCID iD icon 1,2; Fluhrer, Regina
1 Institut für Biologische Grenzflächen (IBG), Karlsruher Institut für Technologie (KIT)
2 Institut für Organische Chemie (IOC), Karlsruher Institut für Technologie (KIT)


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.

Verlagsausgabe §
DOI: 10.5445/IR/1000128322
Veröffentlicht am 11.01.2021
DOI: 10.1016/j.isci.2020.101775
Zitationen: 16
Web of Science
Zitationen: 15
Zitationen: 17
Cover der Publikation
Zugehörige Institution(en) am KIT Institut für Biologische Grenzflächen (IBG)
Institut für Organische Chemie (IOC)
Publikationstyp Zeitschriftenaufsatz
Publikationsmonat/-jahr 12.2020
Sprache Englisch
Identifikator ISSN: 2589-0042
KITopen-ID: 1000128322
HGF-Programm 47.02.04 (POF III, LK 01) Zellpopul.auf Biofunk.Oberflächen IBG-4
Erschienen in iScience
Verlag Cell Press
Band 23
Heft 12
Seiten Article: 101775
Vorab online veröffentlicht am 05.11.2020
Nachgewiesen in Dimensions
Web of Science
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