Isolation of circumferential microtubules from platelets without
simultaneous fixation
JG White, E Radha and M Krumwiede
Circumferential bands of microtubules (MT) support the discoid shape of
resting platelets and participate with the contractile apparatus in shape
change and internal contraction following activation. Elucidation of
interactions between the circumferential coils and proteins of the stable
and contractile cytoskeleton is essential for understanding MT function in
platelet physiology. A previous investigation demonstrated that the
circumferential rings can be isolated intact from resting platelets
following simultaneous exposure to glutaraldehyde and Triton X-100.
However, the use of fixation prevented the characterization of protein
interactions. The present study has circumvented this problem by developing
a procedure for isolating intact microtubule coils from detergent-treated
platelets without the use of fixative agents. Incubation of the platelets
for intervals of 30 to 60 minutes with the microtubule-stabilizing agent
taxol preserved the circumferential bundle after extraction with Triton
X-100 even after washing five times. The procedure has made it possible to
carry out protein studies on isolated microtubule rings and associated
proteins.
Volume 67,
Issue 4,
pp. 873-877,
04/01/1986
Copyright © 1986 by The American Society of Hematology
