Gel-filtered human platelets. Ultrastructure, function, and role of proteins in inhibition of aggregation by aspirin.
Gel filtration of human platelet-rich plasma (PRP) on columns of Sepharose 2B removed at least 99.85% of the plasma proteins from platelets when a column 10 cm in height was used and a plasma volume 11 to 14% of the gel-bed volume was applied. ADP and ATP levels in gel-filtered platelets (GFP) were not significantly different from those in PRP. By transmission electron microscopy, GFP were indistinguishable from PRP. Gel filtration appears to be a highly satisfactory technique of separating platelets from plasma without modifying structure, function, or contents significantly. The roles of several crude protein fractions in platelet aggregation and aspirin's inhibition of aggregation were examined. Fraction I (mostly fibrinogen) enhanced collagen-induced aggregation of gel-filtered platelets; Fraction V (mostly albumin) was inhibitory. Fraction II (mostly gamma-globulin) or gelatin had no significant effect. Aspirin added to gel-filtered platelets inhibited aggregation by 80%. The addition of mixtures of plasma proteins containing albumin increased albumin's inhibitory effect. Incubation of gel-filtered platelets with aspirin labeled in the carboxyl position resulted in no uptake of the label. In contrast, incubation with acetyl-labeled aspirin was followed by uptake of more than 2 X 10(6) acetyl groups per platelet in 1 minute. Incubation for 30 minutes resulted in a five- to sixfold further increase in uptake of the label. Aspirin can acetylate platelets and inhibit aggregation directly. Plasma proteins, in particular albumin or a contaminant of the albumin fraction tested, enhance the inhibitory effect of aspirin on platelet aggregation.
D000107:Acetylation; D001241:Aspirin / Q000494:pharmacology*; D001792:Blood Platelets / Q000032:analysis / Q000502:physiology* / Q000648:ultrastructure; D001798:Blood Proteins / Q000502:physiology*; D002458:Cell Fractionation; D002850:Chromatography, Gel; D005340:Fibrinogen / Q000494:pharmacology; D005780:Gelatin / Q000494:pharmacology; D006801:Humans; D066298:In Vitro Techniques; D010974:Platelet Aggregation / Q000187:drug effects*; D012709:Serum Albumin / Q000494:pharmacology; D005719:gamma-Globulins / Q000494:pharmacology
D001798:Blood Proteins; D012709:Serum Albumin; D005719:gamma-Globulins; D005780:Gelatin; D005340:Fibrinogen; D001241:Aspirin