May 2008
Volume 49, Issue 13
Free
ARVO Annual Meeting Abstract  |   May 2008
Differential Binding of the 402Y and AMD-Associated 402H Variants of Complement Factor H
Author Affiliations & Notes
  • S. J. Clark
    University of Manchester, Manchester, United Kingdom
    Faculty of Life Sciences,
  • R. B. Sim
    MRC Immunochemistry Unit, Department of Biochemistry, University of Oxford, Oxford, United Kingdom
  • A. J. Day
    University of Manchester, Manchester, United Kingdom
    Faculty of Life Sciences,
  • P. N. Bishop
    University of Manchester, Manchester, United Kingdom
    Medical & Human Sciences,
  • Footnotes
    Commercial Relationships  S.J. Clark, None; R.B. Sim, None; A.J. Day, None; P.N. Bishop, None.
  • Footnotes
    Support  University of Manchester Intellectual Property Investment Committee Proof of Principle fund
Investigative Ophthalmology & Visual Science May 2008, Vol.49, 5159. doi:https://doi.org/
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    • Get Citation

      S. J. Clark, R. B. Sim, A. J. Day, P. N. Bishop; Differential Binding of the 402Y and AMD-Associated 402H Variants of Complement Factor H. Invest. Ophthalmol. Vis. Sci. 2008;49(13):5159. doi: https://doi.org/.

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      © ARVO (1962-2015); The Authors (2016-present)

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Abstract

Purpose: : The Y402H variant of complement factor H (CFH) is strongly associated with AMD. The purpose of this study is to further understanding of the pathogenesis of this condition by investigating the differential binding of the 402H and 402Y forms.

Methods: : Two recombinant proteins composed of the complement control protein modules 6-8 of complement factor H containing the 402Y or the 402H variant were expressed and purified. These were used in plate-based assays with a biotinylated glycosaminoglycan to detect differentially binding molecules. The two recombinant proteins were labeled with fluorophores (402Y with AlexFluor-488 and 402H with AlexaFluor-594), incubated simultaneously with 5 µm thick frozen sections of human macula and binding was analyzed by fluorescence microscopy.

Results: : The two recombinant proteins showed differential binding to the macula sections. The distribution differed around choroidal blood vessels and the 402H form bound less strongly to Bruch’s membrane than the 402Y form. The plate-based assays demonstrated differential binding of a form of heparin.

Conclusions: : The differential localization of the 402H and 402Y CFH forms in the macula tissue and their differential binding to a specific glycosaminoglycan suggest that the two forms bind differentially to certain tissue polyanions. In the case of the 402H form this alters the fine balance of complement regulation resulting in a strong predisposition towards AMD.

Keywords: age-related macular degeneration • immunomodulation/immunoregulation • proteoglycans/glycosaminoglycans 
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