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Thromb Haemost 1982; 48(02): 211-216
DOI: 10.1055/s-0038-1657259
Original Article
Schattauer GmbH Stuttgart

Functional Fractionation of Platelets: Aggregation Kinetics and Glycoprotein Labeling of Differing Platelet Populations

V M Haver
The Department of Biochemistry, University of Virginia School of Medicine, Charlottesville, Virginia, U.S.A.
,
A R L Gear
The Department of Biochemistry, University of Virginia School of Medicine, Charlottesville, Virginia, U.S.A.
› Author Affiliations
Further Information

Publication History

Received 10 February 1982

Accepted 21 August 1982

Publication Date:
13 July 2018 (online)

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Summary

Platelet heterogeneity has been studied with a technique called functional fractionation which employs gentle centrifugation to yield subpopulations (“reactive” and “less-reactive” platelets) after exposure to small doses of aggregating agent. Aggregation kinetics of the different platelet populations were investigated by quenched-flow aggregometry. The large, “reactive” platelets were more sensitive to ADP (Ka = 1.74 μM) than the smaller “less-reactive” platelets (Ka = 4.08 μM). However, their maximal rate of aggregation (Vmax, % of platelets aggregating per sec) of 23.3 was significantly lower than the “less-reactive” platelets (Vmax = 34.7). The “reactive” platelets had a 2.2 fold higher level of cyclic AMP.

Platelet glycoproteins were labeled using the neuraminidase-galactose oxidase – [H3]-NaBH4 technique. When platelets were labeled after reversible aggregation, the “reactive” platelets showed a two-fold decrease in labeling efficiency (versus control platelets). However, examination of whole cells or membrane preparations from reversibly aggregated platelets revealed no significant difference in Coomassie or PAS (Schiff) staining.

These results suggest that the large, “reactive” platelets are more sensitive to ADP but are not hyperaggregable in a kinetic sense. Reversible aggregation may cause a re-orientation of membrane glycoproteins that is apparently not characterized by a major loss of glycoprotein material.

Keywords

Platelets - Functional fractionation - Aggregation kinetics - Membrane glycoproteins
 

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