Morphological changes in diabetic kidney are associated with increased O-GlcNAcylation of cytoskeletal proteins including α-actinin 4

Clinical Proteomics

Hayato Kawakami¹  Tamao Endo³  Gerald W Hart⁶  Geert-Jan Boons²  Lance Wells⁴  Margreet A Wolfert⁵  Tosifusa Toda³  Yuri Miura³  Yoshihiro Akimoto¹ 

[1] Department of Anatomy, Kyorin University School of Medicine, Mitaka, Tokyo 181-8611, Japan;Department of Chemistry, University of Georgia, Athens, GA 30602, USA;Research Team for Mechanism of Aging, Tokyo Metropolitan Institute of Gerontology, Itabashi, Tokyo 173-0015, Japan;Department of Biochemistry and Molecular Biology, University of Georgia, Athens, GA 30602, USA;Complex Carbohydrate Research Center, University of Georgia, Athens, GA 30602, USA;Department of Biological Chemistry, Johns Hopkins University School of Medicine, Baltimore, MD 21205, USA

关键词: Proximity Ligation Assay;    Mass spectrometry;    GK Rat;    α-actinin;    Cytoskeleton;    Glomerulus;    Kidney;    Hexosamine biosynthetic pathway;    O-GlcNAc modification;   

Others  :  1026365
DOI  :  10.1186/1559-0275-8-15

 received in 2011-09-02, accepted in 2011-09-21,  发布年份 2011

【 摘 要 】

Purpose

The objective of the present study is to identify proteins that change in the extent of the modification with O-linked N-acetylglucosamine (O-GlcNAcylation) in the kidney from diabetic model Goto-Kakizaki (GK) rats, and to discuss the relation between O-GlcNAcylation and the pathological condition in diabetes.

Methods

O-GlcNAcylated proteins were identified by two-dimensional gel electrophoresis, immunoblotting and peptide mass fingerprinting. The level of O-GlcNAcylation of these proteins was examined by immunoprecipitation, immunoblotting and in situ Proximity Ligation Assay (PLA).

Results

O-GlcNAcylated proteins that changed significantly in the degree of O-GlcNAcylation were identified as cytoskeletal proteins (α-actin, α-tubulin, α-actinin 4, myosin) and mitochondrial proteins (ATP synthase β, pyruvate carboxylase). The extent of O-GlcNAcylation of the above proteins increased in the diabetic kidney. Immunofluorescence and in situ PLA studies revealed that the levels of O-GlcNAcylation of actin, α-actinin 4 and myosin were significantly increased in the glomerulus and the proximal tubule of the diabetic kidney. Immunoelectron microscopy revealed that immunolabeling of α-actinin 4 is disturbed and increased in the foot process of podocytes of glomerulus and in the microvilli of proximal tubules.

Conclusion

These results suggest that changes in the O-GlcNAcylation of cytoskeletal proteins are closely associated with the morphological changes in the podocyte foot processes in the glomerulus and in microvilli of proximal tubules in the diabetic kidney. This is the first report to show that α-actinin 4 is O-GlcNAcylated. α-Actinin 4 will be a good marker protein to examine the relation between O-GlcNAcylation and diabetic nephropathy.

【 授权许可】

   
2011 Akimoto et al; licensee BioMed Central Ltd.

【 图 表 】

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