Summary
The uptake and binding of antimitotic alkaloid colchicine has been demonstrated in
washed preparations of human platelets. A silicone oil technique was adapted so that
both uptake and binding of 14C-colchicine were examined in the same platelet preparations. The time dependence
and amount of colchicine taken up and bound by different platelet preparations during
a 90 to 120 min incubation period were highly reproducible. Both colchicine uptake
and binding by intact platelets, and colchicine binding by preparations of lysed platelets
were specific and temperature dependent. Colchicine uptake was slowly reversible.
Magnesium and GTP enhanced colchicine binding by lysed platelet preparations but calcium
decreased binding.
Exposure of platelets to either cold (4° C) or to thrombin, which disrupt platelet
microtubules, produced significant increases in colchicine uptake and binding. The
thrombin effect was maximal at 37° C and resulted in a greater increase in uptake
and binding than that produced by either cold treatment alone or, by cold treatment
followed by incubation with thrombin at 37° C. The amount of increase in uptake and
binding produced by thrombin was independent of both thrombin (1–5 Units/109 platelets) and colchicine concentrations (1–50 × 10−6M).It is postulated that thrombin may initiate the formation, or make available, colchicine
binding sites (microtubule subunits) within platelets.
