Proton myo-inositol cotransporter is a novel γ-secretase associated protein that regulates Aβ production without affecting Notch cleavage

Yasuhiro Teranishi; Mitsuhiro Inoue; Natsuko Goto Yamamoto; Takahiro Kihara; Birgitta Wiehager; Taizo Ishikawa; Bengt Winblad; Sophia Schedin-Weiss; Susanne Frykman; Lars O Tjernberg

doi:10.1111/febs.13353

Proton myo-inositol cotransporter is a novel γ-secretase associated protein that regulates Aβ production without affecting Notch cleavage

FEBS J. 2015 Sep;282(17):3438-51. doi: 10.1111/febs.13353. Epub 2015 Jul 14.

Authors

Yasuhiro Teranishi^{1

2}, Mitsuhiro Inoue^{1

2}, Natsuko Goto Yamamoto^{1

2}, Takahiro Kihara^{1

3}, Birgitta Wiehager¹, Taizo Ishikawa⁴, Bengt Winblad¹, Sophia Schedin-Weiss¹, Susanne Frykman¹, Lars O Tjernberg¹

Affiliations

¹ Division for Neurogeriatrics, Department of NVS, Center for Alzheimer Research, Karolinska Institutet, Huddinge, Sweden.
² Dainippon Sumitomo Pharma Co., Ltd, Drug Development Research Laboratories, Osaka, Japan.
³ Dainippon-Sumitomo Pharma Co., Ltd, Genomic Science Laboratories, Osaka, Japan.
⁴ Dainippon Sumitomo Pharma Co., Ltd, Innovative Drug Discovery Research Laboratories, Osaka, Japan.

PMID: 26094765
DOI: 10.1111/febs.13353

Abstract

γ-Secretase is a transmembrane protease complex that is responsible for the processing of a multitude of type 1 transmembrane proteins, including the amyloid precursor protein and Notch. γ-Secretase processing of amyloid precursor protein results in the release of the amyloid β-peptide (Aβ), which is involved in the pathogenesis in Alzheimer's disease. Processing of Notch leads to the release of its intracellular domain, which is important for cell development. γ-Secretase associated proteins (GSAPs) could be of importance for substrate selection, and we have previously shown that affinity purification of γ-secretase in combination with mass spectrometry can be used for finding such proteins. In the present study, we used this methodology to screen for novel GSAPs from human brain, and studied their effect on Aβ production in a comprehensive gene knockdown approach. Silencing of probable phospholipid-transporting ATPase IIA, brain-derived neurotrophic factor/neurotrophin-3 growth factor receptor precursor and proton myo-inositol cotransporter (SLC2A13) showed a clear reduction of Aβ and these proteins were selected for further studies on Aβ production and Notch cleavage using small interfering RNA-mediated gene silencing, as well as an overexpression approach. Silencing of these reduced Aβ secretion in a small interfering RNA dose-dependent manner. Interestingly, SLC2A13 had a lower effect on Notch processing. Furthermore, overexpression of SLC2A13 increased Aβ40 generation. Finally, the interaction between γ-secretase and SLC2A13 was confirmed using immunoprecipitation and a proximity ligation assay. In summary, SLC2A13 was identified as a novel GSAP that regulates Aβ production without affecting Notch cleavage. We suggest that SLC2A13 could be a target for Aβ lowering therapy aimed at treating Alzheimer's disease.

Keywords: Alzheimer's disease; Notch; amyloid β-peptide; proton myo-inositol cotransporter; γ-secretase.

Publication types

Research Support, Non-U.S. Gov't
Review

MeSH terms

Adenosine Triphosphatases / antagonists & inhibitors
Adenosine Triphosphatases / genetics
Adenosine Triphosphatases / metabolism
Amyloid Precursor Protein Secretases / genetics*
Amyloid Precursor Protein Secretases / metabolism
Amyloid beta-Peptides / antagonists & inhibitors
Amyloid beta-Peptides / biosynthesis
Amyloid beta-Peptides / genetics*
Animals
Brain Chemistry
Brain-Derived Neurotrophic Factor / antagonists & inhibitors
Brain-Derived Neurotrophic Factor / genetics
Brain-Derived Neurotrophic Factor / metabolism
Carbamates / pharmacology
Dipeptides / pharmacology
Enzyme Inhibitors / pharmacology
Gene Expression Regulation
Gene Knockdown Techniques
Glucose Transport Proteins, Facilitative / antagonists & inhibitors
Glucose Transport Proteins, Facilitative / genetics*
Glucose Transport Proteins, Facilitative / metabolism
Hippocampus / cytology
Hippocampus / drug effects
Hippocampus / metabolism
Humans
Inositol / metabolism
Mice
Microsomes / chemistry
Microsomes / drug effects
Microsomes / metabolism
Molecular Sequence Annotation
Neurons / cytology
Neurons / drug effects
Neurons / metabolism
Peptide Fragments / antagonists & inhibitors
Peptide Fragments / biosynthesis
Peptide Fragments / genetics*
Phospholipid Transfer Proteins / antagonists & inhibitors
Phospholipid Transfer Proteins / genetics
Phospholipid Transfer Proteins / metabolism
Primary Cell Culture
Protein Binding
Protein Stability
Proteolysis
Protons*
RNA, Small Interfering / genetics
RNA, Small Interfering / metabolism
Receptors, Notch / genetics*
Receptors, Notch / metabolism
Signal Transduction

Substances

Amyloid beta-Peptides
Brain-Derived Neurotrophic Factor
Carbamates
Dipeptides
Enzyme Inhibitors
Glucose Transport Proteins, Facilitative
L 685458
Peptide Fragments
Phospholipid Transfer Proteins
Protons
RNA, Small Interfering
Receptors, Notch
SLC2A13 protein, human
amyloid beta-protein (1-40)
Inositol
Amyloid Precursor Protein Secretases
Adenosine Triphosphatases
Atp8a1 protein, mouse