Comparative Proteomic Analysis of the Mitochondria-associated ER Membrane (MAM) in a Long-term Type 2 Diabetic Rodent Model

Scientific Reports
Jacey Hongjie MaSarah X Zhang

Abstract

The mitochondria-associated ER membrane (MAM) plays a critical role in cellular energetics and calcium homeostasis; however, how MAM is affected under diabetic condition remains elusive. This study presented a comprehensive proteome profiling of isolated brain MAM from long-term type 2 diabetic mice vs. non-diabetic controls. MAM protein was extracted efficiently by a surfactant-aided precipitation/on-pellet digestion (SOD) method, and MAM proteome was quantified by an ion-current-based MS1 method combined with nanoLC-MS/MS. A total of 1,313 non-redundant proteins of MAM were identified, among which 144 proteins were found significantly altered by diabetes. In-depth IPA analysis identified multiple disease-relevant signaling pathways associated with the MAM proteome changes in diabetes, most significantly the unfolded protein response (UPR), p53, hypoxia-related transcription factors, and methyl CpG binding protein 2. Using immunofluorescence labeling we confirmed the activation of three UPR branches and increased ERp29 and calreticulin in diabetic retinas. Moreover, we found GRP75, a key MAM tethering protein, was drastically reduced by long-term diabetes. In vitro, acute high glucose treatment reduces ER-mitochondrial contact...Continue Reading

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Citations

May 18, 2020·Proceedings of the National Academy of Sciences of the United States of America·Chulhwan KwakHyun-Woo Rhee
Nov 23, 2017·Frontiers in Cell and Developmental Biology·Gonzalo Soto-HerederoMaría Mittelbrunn
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Aug 9, 2020·Frontiers in Cell and Developmental Biology·Ming YangLin Sun
May 12, 2020·Frontiers in Cell and Developmental Biology·Yixun SuChenju Yi
Jan 24, 2021·Molecular Metabolism·Imke L LemmerAlexander Bartelt
Nov 17, 2020·Cellular and Molecular Neurobiology·Reza Raeisossadati, Merari F R Ferrari
Feb 24, 2021·Nature Reviews. Cardiology·Jun RenYingmei Zhang
Mar 7, 2021·Biomedicines·Nuno Santos Leal, Luís Miguel Martins
May 1, 2021·International Journal of Molecular Sciences·Vladimir ZhemkovIlya Bezprozvanny
Jun 23, 2021·The Journal of Experimental Medicine·Peng XuPaul Greengard

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Methods Mentioned

BETA
Fluorescence
confocal microscopy
proteomic profiling

Software Mentioned

NIH
SEQUEST
R
IonStarstat
ImageJ ( JACoP )
Ingenuity Pathway Analysis ( IPA )
Themo
Image J
SIEVE
Proteome

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