Identification of tubular heparan sulfate as a docking platform for the alternative complement component properdin in proteinuric renal disease.
Properdin binds to proximal tubular epithelial cells (PTEC) and activates the complement system via the alternative pathway in vitro. Cellular ligands for properdin in the kidney have not yet been identified. Because properdin interacts with solid-phase heparin, we investigated whether heparan sulfate proteoglycans (HSPG) could be the physiological ligands of properdin. Kidneys from proteinuric rats showed colocalization of syndecan-1, a major epithelial HSPG, and properdin in the apical membranes of PTEC, which was not seen in control renal tissue. In vitro, PTEC did not constitutively express properdin. However, exogenous properdin binds to these cells in a dose-dependent fashion. Properdin binding was prevented by heparitinase pretreatment of the cells and was dose-dependently inhibited by exogenous heparin. ELISA and surface plasmon resonance spectroscopy (BIAcore) showed a strong dose-dependent interaction between heparan sulfate (HS) and properdin (K(d) = 128 nm). Pretreatment of HSPG with heparitinase abolished this interaction in ELISA. Competition assays, using a library of HS-like polysaccharides, showed that sulfation pattern, chain length, and backbone composition determine the interaction of properdin with glycosaminoglycans. Interestingly, two nonanticoagulant heparin derivatives inhibited properdin-HS interaction in ELISA and BIAcore. Incubation of PTEC with human serum as complement source led to complement activation and deposition of C3 on the cells. This C3 deposition is dependent on the binding of properdin to HS as shown by heparitinase pretreatment of the cells. Our data identify tubular HS as a novel docking platform for alternative pathway activation via properdin, which might play a role in proteinuric renal damage. Our study also suggests nonanticoagulant heparinoids may provide renoprotection in complement-dependent renal diseases.
Department of Nephrology, University Medical Center, 9713 GZ Groningen, The Netherlands. a.zaferani@med.umcg.nl
Azadeh
A
Zaferani
Romain R
RR
Vivès
Pieter
P
van der Pol
Jelleke J
JJ
Hakvoort
Gerjan J
GJ
Navis
Harry
H
van Goor
Mohamed R
MR
Daha
Hugues
H
Lortat-Jacob
Marc A
MA
Seelen
Jacob
J
van den Born
D000818:Animals; D003176:Complement C3 / Q000378:metabolism; D003170:Complement Pathway, Alternative / Q000187:drug effects; D004847:Epithelial Cells / Q000378:metabolism* / Q000473:pathology; D019763:Heparin Lyase / Q000494:pharmacology; D006497:Heparan Sulfate / Q000378:metabolism*; D006801:Humans; D007155:Immunologic Factors / Q000378:metabolism / Q000494:pharmacology; D007674:Kidney Diseases / Q000378:metabolism* / Q000473:pathology; D007687:Kidney Tubules, Proximal / Q000378:metabolism* / Q000473:pathology; D008297:Male; D011414:Properdin / Q000378:metabolism* / Q000494:pharmacology; D011485:Protein Binding / Q000187:drug effects; D011507:Proteinuria / Q000378:metabolism*; D051381:Rats; D017208:Rats, Wistar
D003176:Complement C3; D019763:Heparin Lyase; D006497:Heparan Sulfate; D007155:Immunologic Factors; D011414:Properdin