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Sci Rep. 2017 May 18;7(1):2062. doi: 10.1038/s41598-017-02213-1.

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

Ma JH1,2,3, Shen S4,5, Wang JJ1,2, He Z4,6, Poon A4, Li J5,7, Qu J5,7, Zhang SX8,9,10.

Author information

1
Department of Ophthalmology and Ross Eye Institute, University at Buffalo, State University of New York, Buffalo, NY, USA.
2
SUNY Eye Institute, State University of New York, New York, NY, USA.
3
Aier School of Ophthalmology, Central South University, Changsha, China.
4
Department of Biochemistry, University at Buffalo, State University of New York, Buffalo, NY, USA.
5
New York State Center of Excellence in Bioinformatics and Life Sciences, 701 Ellicott Street, Buffalo, NY, USA.
6
Department of Pediatrics, Sun Yat-Sen Memorial Hospital, Sun Yat-sen University, Guangzhou, China.
7
Department of Pharmaceutical Sciences, School of Pharmacy and Pharmaceutical Sciences, University at Buffalo, State University of New York, Buffalo, NY, USA.
8
Department of Ophthalmology and Ross Eye Institute, University at Buffalo, State University of New York, Buffalo, NY, USA. xzhang38@buffalo.edu.
9
SUNY Eye Institute, State University of New York, New York, NY, USA. xzhang38@buffalo.edu.
10
Department of Biochemistry, University at Buffalo, State University of New York, Buffalo, NY, USA. xzhang38@buffalo.edu.

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 in retinal endothelial cells. This study provides first insight into the significant alterations in MAM proteome associated with activation of the UPR in diabetes, which may serve as novel benchmarks for the future studies of diabetic complications.

PMID:
28522876
PMCID:
PMC5437025
DOI:
10.1038/s41598-017-02213-1
[Indexed for MEDLINE]
Free PMC Article

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