Analysis of the Proteolytic Processing of ABCA3 : Identification of Cleavage Site and Involved Proteases

Hofmann, Nicole; Galetskiy, Dmitry; Rauch, Daniela; Wittmann, Thomas; Marquardt, Andreas; Griese, Matthias; Zarbock, Ralf

Analysis of the Proteolytic Processing of ABCA3 : Identification of Cleavage Site and Involved Proteases

Dateien

Hofmann_0-333803.pdfGröße: 4.14 MBDownloads: 332

Datum

2016

Autor:innen

Hofmann, Nicole

Galetskiy, Dmitry

Rauch, Daniela

Wittmann, Thomas

Marquardt, Andreas

Griese, Matthias

Zarbock, Ralf

Publikationstyp

Zeitschriftenartikel

Publikationsstatus

Published

Erschienen in

PLoS ONE. 2016, 11(3), e0152594. eISSN 1932-6203. Available under: doi: 10.1371/journal.pone.0152594

Zusammenfassung

Rationale
ABCA3 is a lipid transporter in the limiting membrane of lamellar bodies in alveolar type II cells. Mutations in the ABCA3 gene cause respiratory distress syndrome in new-borns and childhood interstitial lung disease. ABCA3 is N-terminally cleaved by an as yet unknown protease, a process believed to regulate ABCA3 activity.

Methods
The exact site where ABCA3 is cleaved was localized using mass spectrometry (MS). Proteases involved in ABCA3 processing were identified using small molecule inhibitors and siRNA mediated gene knockdown. Results were verified by in vitro digestion of a synthetic peptide substrate mimicking ABCA3’s cleavage region, followed by MS analysis.

Results
We found that cleavage of ABCA3 occurs after Lys¹⁷⁴ which is located in the proteins’ first luminal loop. Inhibition of cathepsin L and, to a lesser extent, cathepsin B resulted in attenuation of ABCA3 cleavage. Both enzymes showed activity against the ABCA3 peptide in vitro with cathepsin L being more active.

Conclusion
We show here that, like some other proteins of the lysosomal membrane, ABCA3 is a substrate of cathepsin L. Therefore, cathepsin L may represent a potential target to therapeutically influence ABCA3 activity in ABCA3-associated lung disease.

Fachgebiet (DDC)

570 Biowissenschaften, Biologie

Zitieren

ISO 690

HOFMANN, Nicole, Dmitry GALETSKIY, Daniela RAUCH, Thomas WITTMANN, Andreas MARQUARDT, Matthias GRIESE, Ralf ZARBOCK, 2016. Analysis of the Proteolytic Processing of ABCA3 : Identification of Cleavage Site and Involved Proteases. In: PLoS ONE. 2016, 11(3), e0152594. eISSN 1932-6203. Available under: doi: 10.1371/journal.pone.0152594

BibTex

@article{Hofmann2016-03-31Analy-34314,
  year={2016},
  doi={10.1371/journal.pone.0152594},
  title={Analysis of the Proteolytic Processing of ABCA3 : Identification of Cleavage Site and Involved Proteases},
  number={3},
  volume={11},
  journal={PLoS ONE},
  author={Hofmann, Nicole and Galetskiy, Dmitry and Rauch, Daniela and Wittmann, Thomas and Marquardt, Andreas and Griese, Matthias and Zarbock, Ralf},
  note={Article Number: e0152594}
}

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