Abstract
As part of a broader analysis of the mechanism(s) by which the most sensitive (type III) paroxysmal nocturnal hemoglobinuria (PNH) erythrocytes are excessively sensitive to reactive lysis by isolated C5b6, C7, C8, and C9, we have compared type III PNH (PNH-III) and normal human E in respect to both total specific binding of 125I-C9 and the proportion of cell-bound C9 appearing in high molecular weight (HMW) complexes. In a previous report, we found that after exposure to purified C5b6 and 125I-C7, specific C7 binding and, by implication, EC5b-7 formation were equal for PNH-III E and normal E. In the present study, C8-dependent binding of 125I-C9 to PNH-III EC5b-7 and normal EC5b-7 was also similar, although lysis of the PNH-III E was up to five times greater; that is, PNH-III E required fewer bound C9 molecules to produce an effective lytic site than did normal E. To quantify radioactivity in monomeric and HMW forms of membrane-bound C9, lysed and unlysed E were subjected to low ionic strength buffers to convert all E to ghosts. These ghosts were solubilized in 0.1 or 2% SDS (without reduction) and electrophoresed on 2.4-11% polyacrylamide gradient gels followed by autoradiography and densitometric scanning. With 0.1% SDS, broa...Continue Reading
References
Aug 1, 1979·The Journal of Clinical Investigation·C H PackmanJ P Leddy
Aug 1, 1985·Proceedings of the National Academy of Sciences of the United States of America·V W Hu, A Nicholson-Weller
Jun 1, 1985·The Journal of Clinical Investigation·C J ParkerW F Rosse
Apr 1, 1970·The Journal of Experimental Medicine·R A Thompson, P J Lachmann
Apr 1, 1970·The Journal of Experimental Medicine·P J Lachmann, R A Thompson
Jul 1, 1972·Immunochemistry·U R NilssonF B Taylor
Mar 1, 1973·British Journal of Haematology·W F Rosse
Dec 1, 1967·The Journal of Experimental Medicine·M J Polley, H J Müller-Eberhard
Aug 15, 1970·Nature·U K Laemmli
Mar 1, 1966·Immunochemistry·R A NelsonN Tamura
Jun 1, 1983·The Journal of Experimental Medicine·M K PangburnH J Müller-Eberhard
Sep 1, 1983·Proceedings of the National Academy of Sciences of the United States of America·M K PangburnH J Müller-Eberhard
Jul 1, 1984·Molecular Immunology·E R Podack, J Tschopp
Aug 1, 1983·Proceedings of the National Academy of Sciences of the United States of America·A Nicholson-WellerK F Austen
Citations
Sep 15, 1996·Thrombosis Research·J P MaciejewskiE M Sloand
Nov 9, 2007·Nature Biotechnology·Russell P RotherLeonard Bell
Jul 1, 1987·The Journal of Clinical Investigation·G M HänschD Roelcke
Aug 20, 1987·Biochimica Et Biophysica Acta·M K Pangburn, H Walter