PMID- 26728463
OWN - NLM
STAT- MEDLINE
DCOM- 20160803
LR  - 20210205
IS  - 1083-351X (Electronic)
IS  - 0021-9258 (Print)
IS  - 0021-9258 (Linking)
VI  - 291
IP  - 10
DP  - 2016 Mar 4
TI  - Mutations in Complement Factor H Impair Alternative Pathway Regulation on Mouse 
      Glomerular Endothelial Cells in Vitro.
PG  - 4974-81
LID - 10.1074/jbc.M115.702506 [doi]
AB  - Complement factor H (FH) inhibits complement activation and interacts with 
      glomerular endothelium via its complement control protein domains 19 and 20, 
      which also recognize heparan sulfate (HS). Abnormalities in FH are associated 
      with the renal diseases atypical hemolytic uremic syndrome and dense deposit 
      disease and the ocular disease age-related macular degeneration. Although FH 
      systemically controls complement activation, clinical phenotypes selectively 
      manifest in kidneys and eyes, suggesting the presence of tissue-specific 
      determinants of disease development. Recent results imply the importance of 
      tissue-specifically expressed, sulfated glycosaminoglycans (GAGs), like HS, in 
      determining FH binding to and activity on host tissues. Therefore, we 
      investigated which GAGs mediate human FH and recombinant human FH complement 
      control proteins domains 19 and 20 (FH19-20) binding to mouse glomerular 
      endothelial cells (mGEnCs) in ELISA. Furthermore, we evaluated the functional 
      defects of FH19-20 mutants during complement activation by measuring C3b 
      deposition on mGEnCs using flow cytometry. FH and FH19-20 bound dose-dependently 
      to mGEnCs and TNF-alpha treatment increased binding of both proteins, whereas 
      heparinase digestion and competition with heparin/HS inhibited binding. 
      Furthermore, 2-O-, and 6-O-, but not N-desulfation of heparin, significantly 
      increased the inhibitory effect on FH19-20 binding to mGEnCs. Compared with wild 
      type FH19-20, atypical hemolytic uremic syndrome-associated mutants were less 
      able to compete with FH in normal human serum during complement activation on 
      mGEnCs, confirming their potential glomerular pathogenicity. In conclusion, our 
      study shows that FH and FH19-20 binding to glomerular endothelial cells is 
      differentially mediated by HS but not other GAGs. Furthermore, we describe a 
      novel, patient serum-independent competition assay for pathogenicity screening of 
      FH19-20 mutants.
CI  - (c) 2016 by The American Society for Biochemistry and Molecular Biology, Inc.
FAU - Loeven, Markus A
AU  - Loeven MA
AD  - From the Department of Nephrology, Radboud Institute for Molecular Life Sciences, 
      Radboud University Medical Center, 6525 GA Nijmegen, The Netherlands.
FAU - Rops, Angelique L
AU  - Rops AL
AD  - From the Department of Nephrology, Radboud Institute for Molecular Life Sciences, 
      Radboud University Medical Center, 6525 GA Nijmegen, The Netherlands.
FAU - Lehtinen, Markus J
AU  - Lehtinen MJ
AD  - Department of Bacteriology and Immunology, Haartman Institute, University of 
      Helsinki, FIN-00290 Helsinki, Finland.
FAU - van Kuppevelt, Toin H
AU  - van Kuppevelt TH
AD  - Department of Biochemistry, Radboud Institute for Molecular Life Sciences, 
      Radboud University Medical Center, 6500 HB Nijmegen, The Netherlands.
FAU - Daha, Mohamed R
AU  - Daha MR
AD  - Department of Nephrology, Leiden University Medical Center, 2300 RC Leiden, The 
      Netherlands, and.
FAU - Smith, Richard J
AU  - Smith RJ
AD  - Department of Internal Medicine and Otolaryngology, University of Iowa Carver 
      College of Medicine, Iowa City, Iowa 52242.
FAU - Bakker, Marinka
AU  - Bakker M
AD  - From the Department of Nephrology, Radboud Institute for Molecular Life Sciences, 
      Radboud University Medical Center, 6525 GA Nijmegen, The Netherlands.
FAU - Berden, Jo H
AU  - Berden JH
AD  - From the Department of Nephrology, Radboud Institute for Molecular Life Sciences, 
      Radboud University Medical Center, 6525 GA Nijmegen, The Netherlands.
FAU - Rabelink, Ton J
AU  - Rabelink TJ
AD  - Department of Nephrology, Leiden University Medical Center, 2300 RC Leiden, The 
      Netherlands, and.
FAU - Jokiranta, T Sakari
AU  - Jokiranta TS
AD  - Department of Bacteriology and Immunology, Haartman Institute, University of 
      Helsinki, FIN-00290 Helsinki, Finland.
FAU - van der Vlag, Johan
AU  - van der Vlag J
AD  - From the Department of Nephrology, Radboud Institute for Molecular Life Sciences, 
      Radboud University Medical Center, 6525 GA Nijmegen, The Netherlands, 
      johan.vandervlag@radboudumc.nl.
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
DEP - 20160104
PL  - United States
TA  - J Biol Chem
JT  - The Journal of biological chemistry
JID - 2985121R
RN  - 0 (Glycosaminoglycans)
RN  - 0 (Tumor Necrosis Factor-alpha)
RN  - 80295-65-4 (Complement Factor H)
RN  - 9005-49-6 (Heparin)
SB  - IM
MH  - Animals
MH  - Cell Line
MH  - Complement Activation
MH  - Complement Factor H/chemistry/genetics/immunology/*metabolism
MH  - Endothelial Cells/drug effects/*metabolism
MH  - Glycosaminoglycans/metabolism
MH  - Heparin/pharmacology
MH  - Humans
MH  - Kidney Glomerulus/cytology
MH  - Mice
MH  - *Mutation
MH  - Protein Binding
MH  - Protein Structure, Tertiary
MH  - Tumor Necrosis Factor-alpha/pharmacology
PMC - PMC4777835
OTO - NOTNLM
OT  - atypical hemolytic uremic syndrome
OT  - autoimmune disease
OT  - complement Factor H
OT  - complement system
OT  - endothelial glycocalyx
OT  - endothelium
OT  - heparan sulfate
OT  - innate immunity
EDAT- 2016/01/06 06:00
MHDA- 2016/08/04 06:00
PMCR- 2017/03/04
CRDT- 2016/01/06 06:00
PHST- 2015/11/04 00:00 [received]
PHST- 2016/01/06 06:00 [entrez]
PHST- 2016/01/06 06:00 [pubmed]
PHST- 2016/08/04 06:00 [medline]
PHST- 2017/03/04 00:00 [pmc-release]
AID - S0021-9258(20)43139-4 [pii]
AID - M115.702506 [pii]
AID - 10.1074/jbc.M115.702506 [doi]
PST - ppublish
SO  - J Biol Chem. 2016 Mar 4;291(10):4974-81. doi: 10.1074/jbc.M115.702506. Epub 2016 
      Jan 4.