Cell and Tissue Research

, Volume 337, Issue 1, pp 79–89

Comparison of distinct protein isoforms of the receptor for advanced glycation end-products expressed in murine tissues and cell lines

  • Julia V. Gefter
  • Angel L. Shaufl
  • Mitchell P. Fink
  • Russell L. Delude
Regular Article

DOI: 10.1007/s00441-009-0791-0

Cite this article as:
Gefter, J.V., Shaufl, A.L., Fink, M.P. et al. Cell Tissue Res (2009) 337: 79. doi:10.1007/s00441-009-0791-0
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Abstract

The receptor for advanced glycation end-products (RAGE) is thought to be expressed ubiquitously as various protein isoforms. Our objective was to use Northern blotting, immunoblotting, and sensitivity to N-glycanase digestion to survey RAGE isoforms expressed in cell lines and mouse tissues in order to obtain a more comprehensive view of the RAGE expressome. Pulmonary RAGE mRNA (1.4 kb) was smaller than cell-line and tissue RAGE mRNA (6 kb-10 kb). Three anti-RAGE antibodies that recognized three distinct RAGE epitopes were used for protein studies (N-16, H-300, and αES). Lung expressed three predominant protein isoforms with apparent molecular masses of 45.1, 52.6, and 57.4 kDa (N-16/H-300) and four isoforms at 25.0, 46.9, 52.5, and 54.2 kDa (αES). These isoforms were expressed exclusively in lung. Heart, ileum, and kidney expressed a 44.0-kDa isoform (N-16), whereas aorta and pancreas expressed a 53.3-kDa isoform (αES). Each of these isoforms were absent in tissue extracts prepared from RAGE−/− mice. Cell lines expressed a 70.0-kDa isoform, and a subset expressed a 30.0-kDa isoform (αES). Lung RAGE appeared to contain two N-linked glycans. Tissue and cell-line RAGE isoforms were completely insensitive to PNGase F digestion. Thus, numerous RAGE protein isoforms are detectable in tissues and cell lines. Canonical transmembrane and soluble RAGE appear to be expressed solely in lung (N-16/H-300). Non-pulmonary tissues and cell lines, regardless of the source tissue, both express distinct RAGE protein isoforms containing the N-terminal N-16 epitope or the αES RAGE epitope encoded by alternate exon 9, but lacking the H-300 epitope.

Keywords

Receptor for advanced glycation end productsIsoformsNorthern blottingWestern blottingPNGase FMouse

Copyright information

© Springer-Verlag 2009

Authors and Affiliations

  • Julia V. Gefter
    • 1
    • 2
  • Angel L. Shaufl
    • 1
  • Mitchell P. Fink
    • 1
    • 3
  • Russell L. Delude
    • 1
    • 2
    • 4
  1. 1.Department of Critical Care MedicineUniversity of Pittsburgh School of MedicinePittsburghUSA
  2. 2.Department of PathologyUniversity of Pittsburgh School of MedicinePittsburghUSA
  3. 3.Department of SurgeryUniversity of Pittsburgh School of MedicinePittsburghUSA
  4. 4.615 Scaife HallPittsburghUSA