April 2011
Volume 52, Issue 14
Free
ARVO Annual Meeting Abstract  |   April 2011
Functional Analyses Demonstrate that Common Polymorphisms in C3, Factor B and Factor H Collaborate to Determine Systemic Complement Activity and Risk for AMD
Author Affiliations & Notes
  • Claire L. Harris
    Infection, Immunity & Biochemistry, Cardiff University School of Medicine, Cardiff, United Kingdom
  • Meike Heurich
    Infection, Immunity & Biochemistry, Cardiff University School of Medicine, Cardiff, United Kingdom
  • Ruben Martínez-Barricarte
    Centro de Investigaciones Biológicas, Consejo Superior de Investigaciones Científicas, Madrid, Spain
  • Nigel J. Francis
    Infection, Immunity & Biochemistry, Cardiff University School of Medicine, Cardiff, United Kingdom
  • Dawn L. Roberts
    Infection, Immunity & Biochemistry, Cardiff University School of Medicine, Cardiff, United Kingdom
  • Santiago Rodríguez de Córdoba
    Centro de Investigaciones Biológicas, Consejo Superior de Investigaciones Científicas, Madrid, Spain
  • B Paul Morgan
    Infection, Immunity & Biochemistry, Cardiff University School of Medicine, Cardiff, United Kingdom
  • Footnotes
    Commercial Relationships  Claire L. Harris, None; Meike Heurich, None; Ruben Martínez-Barricarte, None; Nigel J. Francis, None; Dawn L. Roberts, None; Santiago Rodríguez de Córdoba, None; B Paul Morgan, None
  • Footnotes
    Support  Medical Research Council UK, grant G0701298; Spanish Ministerio de Educación y Cultura, grant SAF2008-00226
Investigative Ophthalmology & Visual Science April 2011, Vol.52, 2285. doi:
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      Claire L. Harris, Meike Heurich, Ruben Martínez-Barricarte, Nigel J. Francis, Dawn L. Roberts, Santiago Rodríguez de Córdoba, B Paul Morgan; Functional Analyses Demonstrate that Common Polymorphisms in C3, Factor B and Factor H Collaborate to Determine Systemic Complement Activity and Risk for AMD. Invest. Ophthalmol. Vis. Sci. 2011;52(14):2285.

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

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Abstract

Purpose: : Genetic studies have associated common variations in the complement alternative pathway (AP) proteins C3, factor B (fB) and factor H (fH) with age-related macular degeneration (AMD). We demonstrated that the fB32Q variant forms a C3 convertase less efficiently than the fB32R, leading to decreased amplification of the AP, while fH62I is a better cofactor for factor I-mediated inactivation of C3b than fH62V. These observations explain their protective effects. Here, we perform a functional analysis of the C3R102G variation and investigate how common variation in all three complement proteins combine to affect disease risk.

Methods: : C3, fB and fH variants were purified from plasma of homozygous individuals. Surface plasmon resonance was used to study differences in ligand binding, and haemolysis assays were designed to reveal individual and combined functional differences of complement activating and control proteins.

Results: : C3102G activates the AP more efficiently in whole serum and increases target cell lysis. fB binding kinetics and convertase stability were identical, but native and recombinant fH bound better to C3b102R resulting in significantly decreased cofactor activity. Variants were tested in combination; convertase formed from C3102G and fB32R had three-fold lytic activity compared to C3102R and fB32Q and convertase formed from C3102G and controlled by fH62V showed two-fold increased lysis compared to C3102R and fH62I. Combining disease ‘risk’ variants (C3102G, fB32R, fH62V) yielded six-fold higher AP activity compared to ‘protective’ variants (C3102R, fB32Q, fH62I; P<0.0001).

Conclusions: : We demonstrate here the molecular basis for association of C3102R with AMD, and show that AMD ‘risk’ variants collaborate to enhance AP cycling, while ‘protective’ variants reduce cycling. These data introduce the concept of a functional complotype (combination of polymorphisms) defining complement activity in an individual, and influencing susceptibility to AP-driven disease.

Keywords: age-related macular degeneration • inflammation • protein structure/function 
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