JCI - Mutations in factor H reduce binding affinity to C3b and heparin and surface attachment to endothelial cells in hemolytic uremic syndrome

Mutations in factor H reduce binding affinity to C3b and heparin and surface attachment to endothelial cells in hemolytic uremic syndrome

Tamara Manuelian, Jens Hellwage, Seppo Meri, Jessica Caprioli, Marina Noris, Stefan Heinen, Mihaly Jozsi, Hartmut P.H. Neumann, Giuseppe Remuzzi, Peter F. Zipfel

Published in Volume 111, Issue 8

J Clin Invest. 2003; 111(8):1181–1190 doi:10.1172/JCI16651

Abstract | Full text | PDF

Options: View larger image (or click on image)

Figure 1

Identification of mutant factor H protein in serum of HUS patients. (a) Sera from normal individual (control) (lane 1) and HUS patients with the R1210C (F106, lane 2), the R1215G (F34, lane 3), and the E1172Stop mutation (R043, lane 4) were separated by SDS-PAGE and assayed by Western blotting using anti–factor H antiserum. Note the band with 175 kDa depicted by the arrow in serum of patient F106 (lane 2). Serum derived from patient F34 with the R1215G mutation showed normal factor H, and no additional band was detected. In serum from patient with the E1172Stop mutation (R043), a band of higher mobility with a reduced molecular weight is identified. (b) Reactivity of the normal plasma factor H and the 175 band with antisera and mAb’s specific for factor H. Sera from a healthy individual (lanes 1–3) and patient F106 with the R1210C mutation (lanes 4–10) were separated by SDS-PAGE, and after Western blotting, reacted with the indicated polyclonal and monoclonal Ab’s (lanes 1, 4). (c) Western blotting and (d) silver staining of reduced and deglycosylated (N-glycosidase F; GF) control and patient samples.