title: Resolution and analysis of 'native' and 'activated' properdin.
abstract: A rapid and reproducible procedure for the resolution of 'native' and 'activated' forms of properdin (a component of the alternative activation pathway of complement), by gel filtration on the polyvinyl matrix Fractogel TSK HW-55(S), is reported. This fractionation permitted effective screening of samples for conditions that cause activation. Only 'native' properdin was detected in serum, even after activation of the alternative pathway by yeast cell walls. Transformation of 'native' into 'activated' properdin in vitro was produced by freeze-thawing of the protein, but not upon binding to and dissociation from the C3 convertase, C3bBb. Electron microscopy showed that only the 'native' population contained the discrete cyclic structures described previously by Smith, Pangburn, Vogel & Müller-Eberhard [(1984) J. Biol. Chem. 259, 4582-4588]. 'Activated' properdin, which was eluted from the gel-filtration column close to the breakthrough peak, was mainly composed of large amorphous aggregates. We therefore conclude that properdin 'activation' is not a physiological event that occurs in serum on complement activation, but is an artifact of isolation. Fractionation of properdin on Fractogel TSK HW-55(S) has, however, enabled detailed analysis of functional heterogeneity within the 'native' population.
authors.forename: T C
authors.initials: TC
authors.lastname: Farries
authors.forename: J T
authors.initials: JT
authors.lastname: Finch
authors.forename: P J
authors.initials: PJ
authors.lastname: Lachmann
authors.forename: R A
authors.initials: RA
authors.lastname: Harrison
mesh_terms: D002473:Cell Wall / Q000378:metabolism; D002850:Chromatography, Gel; D003179:Complement C3b / Q000378:metabolism; D003170:Complement Pathway, Alternative; D006801:Humans; D008854:Microscopy, Electron; D011414:Properdin / Q000302:isolation & purification* / Q000378:metabolism; D011485:Protein Binding; D012441:Saccharomyces cerevisiae / Q000378:metabolism
chemical_list: D011414:Properdin; D003179:Complement C3b