Preliminary characterization of a polymorphonuclear leukocyte stimulant isolated from alkali-treated collagen.
This study reports the preliminary characterization of a stimulant released from alkali-treated collagen which activates the respiratory burst of PMNs. The supernatant fraction from alkali-treated collagen (SATC) was precipitated with ammonium sulfate, resuspended, and centrifuged through a sucrose gradient (10-30%, W/V). Proteins were detected throughout the gradient but PMN stimulatory activity was found mainly in fractions 1 and 2 (bottom of gradient), indicating a very high molecular weight. When SATC was layered on a denser sucrose gradient (20-85%, W/V), protein was spread over the upper half of the gradient. The peak of PMN stimulatory activity was found in the middle of the gradient (fractions 8 and 9), indicating a molecular weight similar to the 200,000 dalton standard. High pressure liquid chromatography (HPLC) of fractions from this heavier sucrose gradient showed a small peak, followed by a much higher main peak. The small peak, with the highest molecular weight peptide, is responsible for stimulation of the respiratory burst of PMNs (fraction 8 and 9). The source of this stimulant is yet to be determined.
Pfister
R R
RR
Eye Research Laboratories, AMI/Brookwood Medical Center, Birmingham, Alabama.
Haddox
J L
JL
Lam
K W
KW
Lank
K M
KM
D000468:Alkalies / Q000494:pharmacology*; D002499:Centrifugation, Density Gradient; D005591:Chemical Fractionation; D002851:Chromatography, High Pressure Liquid; D003094:Collagen / Q000032:analysis / Q000378:metabolism*; D006801:Humans; D009504:Neutrophils / Q000378:metabolism / Q000502:physiology*; D010101:Oxygen Consumption / Q000187:drug effects; D012965:Sodium Chloride / Q000494:pharmacology; D013268:Stimulation, Chemical
D000468:Alkalies; D012965:Sodium Chloride; D003094:Collagen