In situ cell surface proteomics reveals differentially expressed membrane proteins in retinal pigment epithelial cells during autoimmune uveitis.
Abstract: Retinal pigment epithelium (RPE) builds the outer blood-retinal barrier of the eye and plays an important role in pathogenesis of the sight threatening disease equine recurrent uveitis (ERU). ERU is a spontaneous autoimmune mediated inflammatory disease characterised by the breakdown of the outer blood-retinal barrier and an influx of autoaggressive T-cells into the inner eye. Therefore, identification of molecular mechanisms contributing to changed function of blood-retinal barrier in ERU is important for the understanding of pathophysiology. Cell surface proteins of RPE collected from healthy horses and horses with ERU were captured by in situ biotinylation and analysed with high resolution mass spectrometry coupled to liquid chromatography (LC-MS/MS) to identify differentially expressed proteins. With label free differential proteomics, a total of 27 differently expressed cell surface proteins in diseased RPE could be detected. Significant down-regulation of three very interesting proteins, synaptotagmin 1, basigin and collectrin was verified and further characterised. Conclusions: We applied an innovative and successful method to detect changes in the plasma cell surface proteome of RPE cells in a spontaneous inflammatory eye disease, serving as a valuable model for human autoimmune uveitis. We were able to identify 27 differentially expressed plasma cell membrane proteins, including synaptotagmin 1, basigin and collectrin, which play important roles in cell adhesion, transport and cell communication.
Copyright © 2014 Elsevier B.V. All rights reserved.
Publication Date: 2014-07-02 PubMed ID: 24998980DOI: 10.1016/j.jprot.2014.06.020Google Scholar: Lookup
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- Journal Article
- Research Support
- Non-U.S. Gov't
Summary
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This research examines the changes in cell surface proteins of Retinal Pigment Epithelium (RPE) cells in horses affected by autoimmune uveitis, a sight-threatening disease. A total of 27 differentially expressed proteins were identified, of which three proteins significantly down-regulated were synaptotagmin 1, basigin and collectrin.
Study Background
- The study focuses on autoimmune uveitis, a disease that threatens vision and results in inflammation of the uvea—the middle layer of the eye that contains most of the eye’s blood vessels.
- It is a spontaneous autoimmune inflammatory illness, notable for its breakdown of the outer blood-retinal barrier and influx of autoaggressive T-cells into the inner eye.
- The disease is particularly prevalent in horses, where it is known as Equine Recurrent Uveitis (ERU).
- Understanding the molecular mechanisms that contribute to changing function of the blood-retinal barrier in ERU could help shed light on pathophysiology.
Experimental Approach
- The protein changes on the RPE cell surfaces were studied using in situ biotinylation—a labelling technique for studying protein interactions.
- High resolution mass spectrometry coupled with liquid chromatography was used to identify proteins that were differentially expressed due to the disease’s influence.
Results
- A total of 27 differentially expressed cell surface proteins in diseased RPE cells were identified.
- Three particularly notable proteins were found to be significantly down-regulated: synaptotagmin 1, basigin and collectrin.
- These proteins are essential for various cellular functions including cell adhesion, transport and cell communication.
Conclusion
- The method used in studying the plasma cell surface proteome was found to be innovative and effective.
- This research serves as a valuable model for human autoimmune uveitis, potentially informing future research into the disease’s pathophysiology.
Cite This Article
APA
Uhl PB, Szober CM, Amann B, Alge-Priglinger C, Ueffing M, Hauck SM, Deeg CA.
(2014).
In situ cell surface proteomics reveals differentially expressed membrane proteins in retinal pigment epithelial cells during autoimmune uveitis.
J Proteomics, 109, 50-62.
https://doi.org/10.1016/j.jprot.2014.06.020 Publication
Researcher Affiliations
- Institute of Animal Physiology, Department of Veterinary Sciences, Ludwig-Maximilians-University, Munich, Germany.
- Institute of Animal Physiology, Department of Veterinary Sciences, Ludwig-Maximilians-University, Munich, Germany.
- Institute of Animal Physiology, Department of Veterinary Sciences, Ludwig-Maximilians-University, Munich, Germany.
- Department of Ophthalmology, Ludwig-Maximilians-University, Munich, Germany.
- Research Unit for Protein Science, Helmholtz Zentrum München, German Research Center for Environmental Health GmbH, Neuherberg, Germany; Centre of Ophthalmology, University Medical Centre, Eberhard-Karls-University Tuebingen, Tuebingen, Germany.
- Research Unit for Protein Science, Helmholtz Zentrum München, German Research Center for Environmental Health GmbH, Neuherberg, Germany.
- Institute of Animal Physiology, Department of Veterinary Sciences, Ludwig-Maximilians-University, Munich, Germany. Electronic address: deeg@tiph.vetmed.uni-muenchen.de.
MeSH Terms
- Animals
- Autoimmune Diseases / metabolism
- Autoimmune Diseases / pathology
- Autoimmune Diseases / veterinary
- Chromatography, Liquid
- Eye Proteins / biosynthesis
- Female
- Gene Expression Regulation
- Horse Diseases / metabolism
- Horse Diseases / pathology
- Horses
- Humans
- Male
- Mass Spectrometry
- Proteomics
- Retinal Pigment Epithelium / metabolism
- Retinal Pigment Epithelium / pathology
- Uveitis / metabolism
- Uveitis / pathology
- Uveitis / veterinary
Citations
This article has been cited 8 times.- Jang HY, Cho CS, Shin YM, Kwak J, Sung YH, Kang BC, Kim JH. Isolation and Characterization of the Primary Marmoset (Callithrix jacchus) Retinal Pigment Epithelial Cells. Cells 2023 Jun 16;12(12).
- Xu S, Xu X, Wu R. Deciphering the Properties and Functions of Glycoproteins Using Quantitative Proteomics. J Proteome Res 2023 Jun 2;22(6):1571-1588.
- Lorenz L, Hirmer S, Schmalen A, Hauck SM, Deeg CA. Cell Surface Profiling of Retinal Müller Glial Cells Reveals Association to Immune Pathways after LPS Stimulation. Cells 2021 Mar 23;10(3).
- Degroote RL, Deeg CA. Immunological Insights in Equine Recurrent Uveitis. Front Immunol 2020;11:609855.
- Shao X, Guha S, Lu W, Campagno KE, Beckel JM, Mills JA, Yang W, Mitchell CH. Polarized Cytokine Release Triggered by P2X7 Receptor from Retinal Pigmented Epithelial Cells Dependent on Calcium Influx. Cells 2020 Nov 24;9(12).
- Fronk AH, Vargis E. Methods for culturing retinal pigment epithelial cells: a review of current protocols and future recommendations. J Tissue Eng 2016 Jan-Dec;7:2041731416650838.
- Grosche A, Hauser A, Lepper MF, Mayo R, von Toerne C, Merl-Pham J, Hauck SM. The Proteome of Native Adult Müller Glial Cells From Murine Retina. Mol Cell Proteomics 2016 Feb;15(2):462-80.
- Uhl PB, Amann B, Hauck SM, Deeg CA. Novel localization of peripherin 2, the photoreceptor-specific retinal degeneration slow protein, in retinal pigment epithelium. Int J Mol Sci 2015 Jan 26;16(2):2678-92.
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