Determination of Complement Factor H Functional Polymorphisms (V62I, Y402H, and E936D) using Sequence-Specific Primer PC
- PDF / 672,141 Bytes
- 8 Pages / 592.44 x 751.18 pts Page_size
- 83 Downloads / 156 Views
ORIGINAL RESEARCH ARTICLE
© 2006 Adis Data Information BV. All rights reserved.
Determination of Complement Factor H Functional Polymorphisms (V62I, Y402H, and E936D) using Sequence-Specific Primer PCR and Restriction Fragment Length Polymorphisms ¨ and Bernadett Blask´o Adrienn B´ir´o, Zolt´an Proh´aszka, George Fust Third Department of Internal Medicine, Faculty of Medicine, Semmelweis University, Szent´agothai J´anos Knowledge Center, Budapest, Hungary
Abstract
Background: Complement factor H (CFH; HF) is an essential regulatory protein that plays a critical role in the homeostasis of the complement system in plasma. Several polymorphisms and mutations in the complement factor H gene (CFH; HF1) have been identified. These have revealed interesting associations with hemolytic-uremic syndrome and age-related macular degeneration. Methods and Results: The aim of this study was to develop a rapid and reliable assay for determining genotypic variants of the CFH gene. Sequence-specific primer PCR and restriction fragment length polymorphism techniques were chosen for the analysis of CFH polymorphisms. The assays detected the following published single nucleotide polymorphisms of CFH in our Caucasian population (n = 271): rs800292, 257G→A (V62I); rs1061170, 1277T→C (Y402H); and rs1065489, 2881G→T (E936D). The allele frequencies (257G = 0.850, 1277T = 0.574, and 2881G = 0.839) that we obtained from a healthy Hungarian population were consistent with previously published results. Conclusion: These analytical methods are simple, reliable, and rapid to perform, and are amenable to automation. Therefore, they could facilitate large-scale genotypic analyses of the CFH gene in various diseases, such as hemolytic-uremic syndrome, age-related macular degeneration, and cardiovascular diseases.
Background The multifunctional and multidomain protein complement factor H (CFH) acts as a central regulator of the complement system and thus represents an essential part of innate immunity.[1] CFH is a 150 kDa single-chain plasma glycoprotein composed of 20 separately folded protein domains, termed ‘short consensus repeats’ (SCRs).[2] It is an abundant humoral regulatory protein involved in the alternative pathway of the complement system. CFH binds to C3b (the activation fragment of the complement protein C3) and accelerates the decay of the alternative C3-convertase (C3bBb) pathway by binding to factor I (cofactor activity).[3,4] CFH has three C3b-binding sites localized to SCR1–4,
SCR12–14, and SCR19–20, respectively.[5] Furthermore, CFH is able to bind to the C-reactive protein heparin and a wide range of pathogen-derived molecules, allowing bacteria to escape complement damage.[1] CFH is encoded by a single gene (CFH; HF1) located on human chromosome 1q32, within the regulators of complement activation (RCA) gene cluster.[6,7] Five additional CFH-related (CFHR) human plasma proteins have been identified and shown to be encoded by five genes (CFHR1–5), which are closely linked to the CFH gene.[8-10] Sequence analyses of the CFH
Data Loading...
