Evidence for the existence of multiple heparan sulfate proteoglycans in the human glomerular basement membrane and mesangial matrix

Alexander J A Groffen, Frank W H Hop, Karl Tryggvason, Henri Dijkman, Karel J M Assmann, Jacques H. Veerkamp, Leo A H Monnens, Lambert P W J Van Den Heuvel

Research output: Contribution to JournalArticleAcademicpeer-review


Heparan sulfate proteoglycans (HSPGs) are essential components of the glomerular basement membrane (GBM) carrying a strong anionic charge. A well- characterized extracellular HSPG is perlecan, ubiquitously expressed in basement membranes. A cDNA construct encoding domains I and II of human perlecan was expressed as a fusion protein with glmathione S-transferase. This fusion protein was used to generate monoclonal antibody 95J10. We compared the staining pattern of 95J10 with that of M215, a previously prepared mAb that recognizes HSPG isolated from human GBM. In kidney cortex, the antiperlecan mAb 95J10 showed a strong staining of the mesangium, Bowman's capsule, the tubular basement membrane, and stained the GBM only slightly. In contrast, M215 predominantly stained the GBM in a linear fashion. Immunoelectron microscopy supported these results, showing concentrations of perlecan in some regions of the GBM, whereas the unidentified M215 antigen was homogemously distributed throughout the GBM. In other human tissues, both antibodies also produced a different staining pattern. Furthermore, a polyclonal antiserum recognizing HSPG isolated from the GBM did not recognize perlecan from EHS tumors. These results provide evidence for the presence of another HSPG in the GBM that is immunologically distinct froth perlecan. The absence of perlecan splice variants in the kidney suggests that this component is encoded by a different gene than perlecan. Given its marked expression in the GBM, this component could be a determining factor in the maintenance of selective glomerular permeability.

Original languageEnglish
Pages (from-to)175-182
Number of pages8
JournalEuropean Journal of Biochemistry
Issue number1
Publication statusPublished - 1997


  • Glomerular basement membrane
  • Heparan sulfate proteoglycan
  • Perlecan
  • Prokaryotic expression


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