Student or educator? Sign up with your school email and get 6 months of Pro free.
CoChat
Sign Up
Cited by 41

The DOI matches its Crossref record; 47 of the 47 references it cites were found in a registry or an index.

ATP6AP2/(Pro)renin Receptor Contributes to Glucose Metabolism via Stabilizing the Pyruvate Dehydrogenase E1 β Subunit

Kanda, Atsuhiro, Noda, Kousuke, Ishida, Susumu·2015·Journal of Biological Chemistry
DOI 10.1074/jbc.m114.626713
Open this paper in CoChat — free account

Abstract

Background: The mechanism of efficient energy generation in the highly evolved mammalian retina remains incompletely understood. Results: ATP6PA2 interacts with the E1 β subunit of pyruvate dehydrogenase, a key enzyme in aerobic energy generation, controlling its protein stability. Conclusion: ATP6AP2 contributes to glucose metabolism together with oxidative stress. Significance: The present data provide a novel insight into the biological function of ATP6AP2 in the retina. Aerobic glucose metabolism is indispensable for metabolically active cells; however, the regulatory mechanism of efficient energy generation in the highly evolved mammalian retina remains incompletely understood. Here, we revealed an unsuspected role for (pro)renin receptor, also known as ATP6AP2, in energy metabolism. Immunoprecipitation and mass spectrometry analyses identified the pyruvate dehydrogenase (PDH) complex as Atp6ap2-interacting proteins in the mouse retina. Yeast two-hybrid assays demonstrated direct molecular binding between ATP6AP2 and the PDH E1 β subunit (PDHB). Pdhb immunoreactivity co-localized with Atp6ap2 in multiple retinal layers including the retinal pigment epithelium (RPE). ATP6AP2 knockdown in RPE cells reduced PDH activity, showing a predilection to anaerobic glycolysis. ATP6AP2 protected PDHB from phosphorylation, thus controlling its protein stability. Down-regulated PDH activity due to ATP6AP2 knockdown inhibited glucose-stimulated oxidative stress in RPE cells. Our present data unraveled the novel function of ATP6AP2/(P)RR as a PDHB stabilizer, contributing to aerobic glucose metabolism together with oxidative stress.

Overview

The claims and statistics of this paper haven’t been checked yet.

Run the check — free, no account needed. It reads the open-access text and checks the sources it cites.

A few free checks a day · papers already verified open instantly · results are public

Integrity signals

Registry

Crossref · 10.1074/jbc.m114.626713

Cites

47 of 47 references verified0 citation errors · 0 not checkable

Statistics

Figures

FIGURE 1 Direct molecular binding of ATP6AP2 with PDHB. A , mouse retina homogenate was applied to anti-ATP6AP2 antibody- or normal IgG-immobilized protein G beads. The eluted proteins were separated by SDS-PAGE and visualized with silver staining. Two protein bands around 35 and 65 kDa were detected ( arrows A and B ). B , known-bait and known-prey yeast twohybrid analyses. Upperpanels represent the growth of yeast transformed with ATP6AP2 (bait) and PDHX , PDHA1 , PDHB , DLD or DLAT (prey) constructs (growth assay). Right panels represent 𝛃 -galactosidase color ( blue ) tests for the growth assay shown in the left panels . Bottompanels represent a bait-prey switch experiment (PDHB, bait; ATP6AP2, prey) together with negative (ATP6AP2 + Empty, Lam (Laminin) + T-Ag (antigen)) and positive (p53 + T-Ag) controls. C , co-IP of mouse retinal extracts using anti-Atp6ap2 antibody was performed followed by SDS-PAGE and immunoblot ( IB ) analyses with anti-Pdhb antibody. D , co-transfection and co-IP. FLAG-tagged ATP6AP2 expressionplasmidswereco-transfectedintoHEK293TcellswithMyc-tagged PDHB expression constructs. IP with anti-Myc antibody was followed by detection with anti-FLAG antibody. Bands at 25 kDa indicate immunoglobulin light chains. Schematic representation of the ATP6AP2 constructs used. SP , signal peptide; CS , furin cleavage site; TM , transmembrane domain.
FIGURE 1 Direct molecular binding of ATP6AP2 with PDHB. A , mouse retina homogenate was applied to anti-ATP6AP2 antibody- or normal IgG-immobilized protein G beads. The eluted proteins were separated by SDS-PAGE and visualized with silver staining. Two protein bands around 35 and 65 kDa were detected ( arrows A and B ). B , known-bait and known-prey yeast twohybrid analyses. Upperpanels represent the growth of yeast transformed with ATP6AP2 (bait) and PDHX , PDHA1 , PDHB , DLD or DLAT (prey) constructs (growth assay). Right panels represent 𝛃 -galactosidase color ( blue ) tests for the growth assay shown in the left panels . Bottompanels represent a bait-prey switch experiment (PDHB, bait; ATP6AP2, prey) together with negative (ATP6AP2 + Empty, Lam (Laminin) + T-Ag (antigen)) and positive (p53 + T-Ag) controls. C , co-IP of mouse retinal extracts using anti-Atp6ap2 antibody was performed followed by SDS-PAGE and immunoblot ( IB ) analyses with anti-Pdhb antibody. D , co-transfection and co-IP. FLAG-tagged ATP6AP2 expressionplasmidswereco-transfectedintoHEK293TcellswithMyc-tagged PDHB expression constructs. IP with anti-Myc antibody was followed by detection with anti-FLAG antibody. Bands at 25 kDa indicate immunoglobulin light chains. Schematic representation of the ATP6AP2 constructs used. SP , signal peptide; CS , furin cleavage site; TM , transmembrane domain.CC-BY · From the PDF
FIGURE 2 Co-localization of Atp6ap2 with Pdhb in the retina. A-C , double labeling of Pdhb ( green ) and Atp6ap2 ( red ) together with DAPI ( blue ). A ′ -C ′ and A ″ -C ″ , images show higher magnification. ONL , outer nuclear layer; INL , inner nuclear layer; GCL , ganglion cell layer. Scale bar = 30 𝛍 m. D and D ′ , immunogold labeling for Atp6ap2 in the cytosol of RPE cells. Scale bar = 1 𝛍 m. E , mouse retinal homogenate was fractionated to the cytosol and membrane followed by SDS-PAGE and immunoblot analyses using anti-Atp6ap2, anti-Hsp90 (cytosolic marker), and anti-Na + /K + -ATPase 𝛂 1 (plasma membrane marker) antibodies. F , ATP6AP2 immunostaining ( purple ) in human RPE. G , no staining with normal IgG. Scale bar = 15 𝛍 m.
FIGURE 2 Co-localization of Atp6ap2 with Pdhb in the retina. A-C , double labeling of Pdhb ( green ) and Atp6ap2 ( red ) together with DAPI ( blue ). A ′ -C ′ and A ″ -C ″ , images show higher magnification. ONL , outer nuclear layer; INL , inner nuclear layer; GCL , ganglion cell layer. Scale bar = 30 𝛍 m. D and D ′ , immunogold labeling for Atp6ap2 in the cytosol of RPE cells. Scale bar = 1 𝛍 m. E , mouse retinal homogenate was fractionated to the cytosol and membrane followed by SDS-PAGE and immunoblot analyses using anti-Atp6ap2, anti-Hsp90 (cytosolic marker), and anti-Na + /K + -ATPase 𝛂 1 (plasma membrane marker) antibodies. F , ATP6AP2 immunostaining ( purple ) in human RPE. G , no staining with normal IgG. Scale bar = 15 𝛍 m.CC-BY · From the PDF
Image 3
CC-BY · From the PDF
Image 4
CC-BY · From the PDF
FIGURE 4 Reduced PDHB protein, but not mRNA, levels and enhanced PDHB phosphorylation following ATP6AP2 knockdown. A , immunoblot ( IB ) analyses for ATP6AP2, PDHA1, DLAT, PDHB, and 𝛃 -actin proteins in ATP6AP2 -knockdownRPEcells.Relative intensity = 0.49 ± 0.12 for siRNA-1, and 0.60 ± 0.18 for siRNA-2, compared with control ( n = 3). B , gene expression levels of PDHB in ATP6AP2 -knockdown RPE cells. Real-time PCR amplification of PDHB after transfection with ATP6AP2 or negative control siRNA (fold-change = 0.97 for siRNA-1 and 1.05 for siRNA-2). C , co-IP of ATP6AP2 -knockdown RPE cell extracts using anti-PDHB antibody followed by SDS-PAGE and IB analyses using anti-PDHB and anti-phosphotyrosine ( p-tyrosine ) antibodies. Relative intensity = 1.24 ± 0.10* for siRNA-1, and 1.25 ± 0.21* for siRNA-2, compared with control (*, p < 0.05, n = 4). D , IB analyses using anti-PDHB antibody following treatment with or without the tyrosine kinase inhibitor AG18 for ATP6AP2 -knockdown RPE cells. Relative intensity = 0.55 ± 0.12* for siRNA-1, 0.65 ± 0.08* for siRNA-2, 0.98 ± 0.14 for control + AG18, 0.94 ± 0.09 for siRNA-1 + AG18, and 0.99 ± 0.05 for siRNA-2 + AG18, compared with control (*, p < 0.05, n = 4). Densitometry values of PDHB and tyrosine-phosphorylated PDHB ( p-PDHB ) normalized to 𝛃 -actin and total PDHB ( t-PDHB ), respectively, are shown in parentheses ( A, C , and D ).
FIGURE 4 Reduced PDHB protein, but not mRNA, levels and enhanced PDHB phosphorylation following ATP6AP2 knockdown. A , immunoblot ( IB ) analyses for ATP6AP2, PDHA1, DLAT, PDHB, and 𝛃 -actin proteins in ATP6AP2 -knockdownRPEcells.Relative intensity = 0.49 ± 0.12 for siRNA-1, and 0.60 ± 0.18 for siRNA-2, compared with control ( n = 3). B , gene expression levels of PDHB in ATP6AP2 -knockdown RPE cells. Real-time PCR amplification of PDHB after transfection with ATP6AP2 or negative control siRNA (fold-change = 0.97 for siRNA-1 and 1.05 for siRNA-2). C , co-IP of ATP6AP2 -knockdown RPE cell extracts using anti-PDHB antibody followed by SDS-PAGE and IB analyses using anti-PDHB and anti-phosphotyrosine ( p-tyrosine ) antibodies. Relative intensity = 1.24 ± 0.10* for siRNA-1, and 1.25 ± 0.21* for siRNA-2, compared with control (*, p < 0.05, n = 4). D , IB analyses using anti-PDHB antibody following treatment with or without the tyrosine kinase inhibitor AG18 for ATP6AP2 -knockdown RPE cells. Relative intensity = 0.55 ± 0.12* for siRNA-1, 0.65 ± 0.08* for siRNA-2, 0.98 ± 0.14 for control + AG18, 0.94 ± 0.09 for siRNA-1 + AG18, and 0.99 ± 0.05 for siRNA-2 + AG18, compared with control (*, p < 0.05, n = 4). Densitometry values of PDHB and tyrosine-phosphorylated PDHB ( p-PDHB ) normalized to 𝛃 -actin and total PDHB ( t-PDHB ), respectively, are shown in parentheses ( A, C , and D ).CC-BY · From the PDF
FIGURE 5 Suppression of glucose-stimulated oxidative stress following ATP6AP2 knockdown. A , ROS production, elevated by high glucose (control siRNA = 135.2%), was reduced by ATP6AP2 knockdown (siRNA-1 = 113.9%, siRNA-2 = 115.1%). B , no significant changes in ROS generation were detected in ATP6AP2 -knockdown cells in which RAPS- and RAS-related pathways were abrogated by PRRB (control siRNA = 134.4%, siRNA-1 = 112.9%, siRNA-2 = 111.5%)andvalsartan(controlsiRNA = 135.4%,siRNA-1 = 113.4%, siRNA-2 = 114.7%), respectively ( n = 12). HG , high glucose; NG , normal glucose; OC , osmotic control. *, p < 0.05; **, p < 0.01.
FIGURE 5 Suppression of glucose-stimulated oxidative stress following ATP6AP2 knockdown. A , ROS production, elevated by high glucose (control siRNA = 135.2%), was reduced by ATP6AP2 knockdown (siRNA-1 = 113.9%, siRNA-2 = 115.1%). B , no significant changes in ROS generation were detected in ATP6AP2 -knockdown cells in which RAPS- and RAS-related pathways were abrogated by PRRB (control siRNA = 134.4%, siRNA-1 = 112.9%, siRNA-2 = 111.5%)andvalsartan(controlsiRNA = 135.4%,siRNA-1 = 113.4%, siRNA-2 = 114.7%), respectively ( n = 12). HG , high glucose; NG , normal glucose; OC , osmotic control. *, p < 0.05; **, p < 0.01.CC-BY · From the PDF
FIGURE 6 A scheme showing ATP6AP2 involvement in PDH-mediated aerobic glucose metabolism and oxidative stress. ATP6AP2 blockade causes a metabolic shift from aerobic cellular respiration to anaerobic glycolysis ( green arrows ), leading consequently to a decrease in mitochondrial ROS.
FIGURE 6 A scheme showing ATP6AP2 involvement in PDH-mediated aerobic glucose metabolism and oxidative stress. ATP6AP2 blockade causes a metabolic shift from aerobic cellular respiration to anaerobic glycolysis ( green arrows ), leading consequently to a decrease in mitochondrial ROS.CC-BY · From the PDF

Verification

Claims the paper makes

The claims and statistics of this paper haven’t been checked yet.

Run the check — free, no account needed. It reads the open-access text and checks the sources it cites.

A few free checks a day · papers already verified open instantly · results are public

Claims other papers make about this one

Each is checked against this paper’s own text: does it say what the citing paper says it does?

Self-check

Percentages, sums and reported differences re-computed from the paper’s own numbers.

The self-check needs the paper’s full text.

Citations

Cites

Show all 47 references

Cited by

See all 41

Metadata

Authors
Kanda, A., Noda, K., & Ishida, S.
Venue
Journal of Biological Chemistry
Publisher
Elsevier BV
Published
2015-04
Volume
290
Issue
15
Pages
9690-9700
ISSN
0021-9258
DOI
10.1074/jbc.m114.626713
Cited by
41
References
47
Language
en
Licence

About these checks

Registry, retraction, Cites and statistics results are computed, not judged, with one exception: a printed reference that no index lists is read once by a model before it counts as a citation error. Claims are extracted by a model and kept only when their quote appears verbatim in the text; each cited source is then checked, with a model’s help, for whether it says what the claim relies on. Those verdicts can be wrong — read the claim’s quote and the cited source before relying on one.

Statistics and arithmetic are recomputed, not judged.

Confidence bands are shown only for fields calibrated on our annotated test set. None is calibrated yet, so no band is shown.

What counts as a claim ›How we measure confidence ›How we check this ›

Are you an author, or do you see a problem with this page? Request a review

How to cite

Cite CoChat for the checks on this page, and the paper for its own findings.

This page
CoChat. (2026). Citation check: ATP6AP2/(Pro)renin Receptor Contributes to Glucose Metabolism via Stabilizing the Pyruvate Dehydrogenase E1 β Subunit. https://app.cochat.ai/papers/10.1074/jbc.m114.626713
The paper
Kanda, A., Noda, K., & Ishida, S. (2015). ATP6AP2/(Pro)renin Receptor Contributes to Glucose Metabolism via Stabilizing the Pyruvate Dehydrogenase E1 β Subunit. Journal of Biological Chemistry. https://doi.org/10.1074/jbc.m114.626713

Data sources

Bibliographic data: Semantic Scholar, OpenAlex, Crossref.

Contains information from the Semantic Scholar Academic Graph Datasets, made available under the ODC Attribution License (ODC-BY 1.0).

Semantic Scholar’s data is described in: Kinney, R., et al. (2023). The Semantic Scholar Open Data Platform. arXiv:2301.10140.