Skip to main content
bioRxiv
  • Home
  • About
  • Submit
  • ALERTS / RSS
Advanced Search
New Results

Stabilization / destabilization of the APP transmembrane domain by mutations in the di-glycine hinge alter helical structure and dynamics, and impair cleavage by γ-secretase

View ORCID ProfileAlexander Götz, Nadine Mylonas, View ORCID ProfilePhilipp Högel, Mara Silber, Hannes Heinel, Simon Menig, Alexander Vogel, Hannes Feyrer, View ORCID ProfileDaniel Huster, Burkhard Luy, View ORCID ProfileDieter Langosch, View ORCID ProfileChristina Scharnagl, Claudia Muhle-Goll, Frits Kamp, Harald Steiner
doi: https://doi.org/10.1101/375006
Alexander Götz
1Physics of Synthetic Biological Systems (E14), Technical University of Munich, Freising, Germany
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
  • ORCID record for Alexander Götz
Nadine Mylonas
2Biomedical Center - BMC, Metabolic Biochemistry, Ludwig-Maximilians-University,Munich, Germany
5German Center for Neurodegenerative Diseases (DZNE), Munich, Germany
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Philipp Högel
3Center for Integrated Protein Science Munich (CIPSM) at the Lehrstuhl Chemie der Biopolymere, Technical University Munich, Weihenstephaner Berg 3, 85354 Freising, Germany
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
  • ORCID record for Philipp Högel
Mara Silber
4Institute of Organic Chemistry and Institute for Biological Interfaces 4, Karlsruhe Institute of Technology, Karlsruhe, Germany
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Hannes Heinel
6Institute for Medical Physics and Biophysics, Leipzig University, Leipzig, Germany
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Simon Menig
1Physics of Synthetic Biological Systems (E14), Technical University of Munich, Freising, Germany
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Alexander Vogel
6Institute for Medical Physics and Biophysics, Leipzig University, Leipzig, Germany
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Hannes Feyrer
4Institute of Organic Chemistry and Institute for Biological Interfaces 4, Karlsruhe Institute of Technology, Karlsruhe, Germany
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Daniel Huster
6Institute for Medical Physics and Biophysics, Leipzig University, Leipzig, Germany
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
  • ORCID record for Daniel Huster
Burkhard Luy
4Institute of Organic Chemistry and Institute for Biological Interfaces 4, Karlsruhe Institute of Technology, Karlsruhe, Germany
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Dieter Langosch
3Center for Integrated Protein Science Munich (CIPSM) at the Lehrstuhl Chemie der Biopolymere, Technical University Munich, Weihenstephaner Berg 3, 85354 Freising, Germany
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
  • ORCID record for Dieter Langosch
Christina Scharnagl
1Physics of Synthetic Biological Systems (E14), Technical University of Munich, Freising, Germany
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
  • ORCID record for Christina Scharnagl
  • For correspondence: christina.scharnagl@tum.de claudia.muhle-goll@kit.edu harald.steiner@med.uni-muenchen.de
Claudia Muhle-Goll
4Institute of Organic Chemistry and Institute for Biological Interfaces 4, Karlsruhe Institute of Technology, Karlsruhe, Germany
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
  • For correspondence: christina.scharnagl@tum.de claudia.muhle-goll@kit.edu harald.steiner@med.uni-muenchen.de
Frits Kamp
2Biomedical Center - BMC, Metabolic Biochemistry, Ludwig-Maximilians-University,Munich, Germany
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Harald Steiner
2Biomedical Center - BMC, Metabolic Biochemistry, Ludwig-Maximilians-University,Munich, Germany
5German Center for Neurodegenerative Diseases (DZNE), Munich, Germany
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
  • For correspondence: christina.scharnagl@tum.de claudia.muhle-goll@kit.edu harald.steiner@med.uni-muenchen.de
  • Abstract
  • Full Text
  • Info/History
  • Metrics
  • Supplementary material
  • Preview PDF
Loading

Abstract

γ-Secretase is a pivotal intramembrane protease that is involved in the generation of toxic Alzheimer´s disease-associated peptides from the γ-amyloid precursor protein C-terminal fragment C99. Determinants of γ-secretase substrates are not defined yet and in the absence of recognition motifs, helix-stability and dynamics have been discussed as critical factors for recognition and cleavage of γ-secretase. Since conformational flexibility of a di-glycine hinge region in the C99 transmembrane domain (TMD), which separates the TMD into N- and C-terminal parts, might play a role in the processing of C99 by γ-secretase, we mutated one of the glycines, G38, to a helix-stabilizing leucine and to a helix-distorting proline residue. While γ-secretase cleavage of the G38L mutant was reduced, that of G38P was strongly impaired. Furthermore, cleavage precision of γ-secretase was dramatically altered by the mutations. CD and NMR spectroscopy, hydrogen/deuterium exchange measurements as well as in silico modeling by MD simulations to assess structural and dynamical parameters showed that the mutations affected the TMD helix properties, in particularly for the G38P mutant. However, helix destabilization was not observed at the #-cleavage sites of C99, suggesting that local unfolding of the TMD to allow access to the scissile bonds requires the presence of the protease. Our data suggest that conformational relaxations of substrate and enzyme during substrate recruitment required for catalysis are established only when substrate and protease come into contact.

Copyright 
The copyright holder for this preprint is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. All rights reserved. No reuse allowed without permission.
Back to top
PreviousNext
Posted July 23, 2018.
Download PDF

Supplementary Material

Email

Thank you for your interest in spreading the word about bioRxiv.

NOTE: Your email address is requested solely to identify you as the sender of this article.

Enter multiple addresses on separate lines or separate them with commas.
Stabilization / destabilization of the APP transmembrane domain by mutations in the di-glycine hinge alter helical structure and dynamics, and impair cleavage by γ-secretase
(Your Name) has forwarded a page to you from bioRxiv
(Your Name) thought you would like to see this page from the bioRxiv website.
CAPTCHA
This question is for testing whether or not you are a human visitor and to prevent automated spam submissions.
Share
Stabilization / destabilization of the APP transmembrane domain by mutations in the di-glycine hinge alter helical structure and dynamics, and impair cleavage by γ-secretase
Alexander Götz, Nadine Mylonas, Philipp Högel, Mara Silber, Hannes Heinel, Simon Menig, Alexander Vogel, Hannes Feyrer, Daniel Huster, Burkhard Luy, Dieter Langosch, Christina Scharnagl, Claudia Muhle-Goll, Frits Kamp, Harald Steiner
bioRxiv 375006; doi: https://doi.org/10.1101/375006
Digg logo Reddit logo Twitter logo Facebook logo Google logo LinkedIn logo Mendeley logo
Citation Tools
Stabilization / destabilization of the APP transmembrane domain by mutations in the di-glycine hinge alter helical structure and dynamics, and impair cleavage by γ-secretase
Alexander Götz, Nadine Mylonas, Philipp Högel, Mara Silber, Hannes Heinel, Simon Menig, Alexander Vogel, Hannes Feyrer, Daniel Huster, Burkhard Luy, Dieter Langosch, Christina Scharnagl, Claudia Muhle-Goll, Frits Kamp, Harald Steiner
bioRxiv 375006; doi: https://doi.org/10.1101/375006

Citation Manager Formats

  • BibTeX
  • Bookends
  • EasyBib
  • EndNote (tagged)
  • EndNote 8 (xml)
  • Medlars
  • Mendeley
  • Papers
  • RefWorks Tagged
  • Ref Manager
  • RIS
  • Zotero
  • Tweet Widget
  • Facebook Like
  • Google Plus One

Subject Area

  • Biophysics
Subject Areas
All Articles
  • Animal Behavior and Cognition (3482)
  • Biochemistry (7329)
  • Bioengineering (5301)
  • Bioinformatics (20212)
  • Biophysics (9985)
  • Cancer Biology (7706)
  • Cell Biology (11273)
  • Clinical Trials (138)
  • Developmental Biology (6425)
  • Ecology (9923)
  • Epidemiology (2065)
  • Evolutionary Biology (13292)
  • Genetics (9353)
  • Genomics (12559)
  • Immunology (7681)
  • Microbiology (18964)
  • Molecular Biology (7421)
  • Neuroscience (40915)
  • Paleontology (298)
  • Pathology (1226)
  • Pharmacology and Toxicology (2130)
  • Physiology (3145)
  • Plant Biology (6842)
  • Scientific Communication and Education (1271)
  • Synthetic Biology (1893)
  • Systems Biology (5299)
  • Zoology (1086)