Purification and partial characterization of rat factor D

Rat factor D has been purified to homogeneity (10559-fold) from serum by chromatography on CM-Sepharose Fast Flow, phenyl-Sepharose CL-4B and Mono S and has been shown to resemble its human and mouse counterparts both in substrate specificity and in its susceptibility to inhibition by the organophosphorous inhibitor di-isopropyl-fluorophosphate. The rat enzyme, however, is heavily glycosylated and binds to wheat-germ lectin-Sepharose 6MB and S-hydroxytryptamine-agarose, but not to concanavalin A-Sepharose 4B. All of the carbohydrate chains are N-linked. Enzymic removal of this carbohydrate decreased the M(r), by approx. 15000. The deglycosylated rat enzyme had the same mobility as native human factor D on SDS/PAGE, corresponding to an M(r), of 24500. N-Terminal sequence analysis of the first 30 amino acids of rat factor D highlighted the sequence similarity with human factor D (> 76%) and, in particular, with mouse adipsin (> 93%).