Purification and characterization of a neutrophil chemotactic factor from Dirofilaria immitis.
A neutrophil chemotactic factor (NCF-Di) was purified from a crude extract of Dirofilaria immitis adult worm by a combination of anion-exchange chromatography on DE52 and gel filtration on Sephacryl S-200. NCF-Di showed a single protein band by both polyacrylamide gel electrophoresis (PAGE) and sodium dodecyl sulfate (SDS) PAGE. The molecular weight of NCF-Di was estimated to be 17,000 by gel filtration on Sephadex G-150, and 14,000 by SDS-PAGE. NCF-Di was an acidic protein with isoelectric point of 4.5. NCF-Di was absorbed neither to lentil lectin-Sepharose nor to concanavalin A-Sepharose. The chemotactic activity of NCF-Di was heat labile (56 C, 1 hr), but was resistant to periodate oxidation. These results suggest that NCF-Di is a simple peptide which has few or no sugar chains. These physicochemical properties of NCF-Di were compared to previously reported parasite-derived chemoattractants or purified allergen of D. immitis.
D000818:Animals; D055598:Chemical Phenomena; D002627:Chemistry, Physical; D002630:Chemotactic Factors / Q000302:isolation & purification*; D002634:Chemotaxis, Leukocyte; D002850:Chromatography, Gel; D002852:Chromatography, Ion Exchange; D004183:Dirofilaria immitis / Q000032:analysis* / Q000469:parasitology; D004285:Dogs; D004591:Electrophoresis, Polyacrylamide Gel; D005370:Filarioidea / Q000032:analysis*; D016209:Interleukin-8; D009504:Neutrophils / Q000469:parasitology / Q000502:physiology*
D002630:Chemotactic Factors; D016209:Interleukin-8