Native polymeric forms of properdin selectively bind to targets and promote activation of the alternative pathway of complement.
Properdin, a positive regulator of the complement system, has recently been reported to bind to certain pathogenic microorganisms, to early or late apoptotic and necrotic cells, and to particular live human cell lines, thus providing a platform for de novo convertase assembly and complement activation. These studies, with some contradictory results, have been carried out with purified properdin, which forms a series of oligomers of a ∼53,000 Mr subunit, assembling into dimers (P₂), trimers (P₃), tetramers (P₄) and higher forms (P(n)). The P(n) forms have been shown to likely be an artefact of purification that results from procedures including freeze-thawing of properdin. In this study we isolated the individual natural forms of properdin (P₂, P₃, and P₄) and separated them from the P(n) forms present in purified frozen properdin using ion exchange and/or size exclusion chromatography. We analyzed the ability of each form to bind to live or necrotic Jurkat and Raji cells, rabbit erythrocytes (E(R)), and zymosan by FACS analysis. While the unseparated properdin and the purified P(n) forms bound to all the surfaces except E(R), the physiological P₂-P₄ forms specifically bound only to zymosan and to necrotic nucleated cells. Our results indicate that aggregated P(n) present in unseparated properdin may bind non-specifically to some surfaces and should be separated before analysis in order to obtain meaningful results. Finally, we have determined for the first time that the physiological forms of human properdin can selectively recognize surfaces and enhance or promote complement activation, which is in agreement with the reported role for properdin as a complement initiator.
Ferreira
Viviana P
VP
Department of Medical Microbiology and Immunology, College of Medicine, University of Toledo, Toledo, OH 43614, USA. viviana.ferreira@utoledo.edu
Cortes
Claudio
C
Pangburn
Michael K
MK
D000818:Animals; D045744:Cell Line, Tumor; D003170:Complement Pathway, Alternative; D004912:Erythrocytes / Q000276:immunology; D006801:Humans; D019169:Jurkat Cells; D011414:Properdin / Q000276:immunology* / Q000302:isolation & purification; D055503:Protein Multimerization; D011817:Rabbits; D015054:Zymosan / Q000276:immunology
D011414:Properdin; D015054:Zymosan
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