Transbilayer incorporation of 1-pyrenebutyltrimethylammonium by blood platelets and its application for analyzing changes in physico-chemical properties of the membrane lipid bilayer induced by platelet activation.
The binding of cationic butyltrimethylammonium derivative of pyrene to bovine platelets was initially rapid and then increased gradually, unlike the bindings of other anionic and neutral derivatives of pyrene tested. The rate of increase in binding of the cationic probe depended on temperature and was due to its incorporation into the cytoplasmic side of the platelet membranes, as shown quantitatively by monitoring decrease in its extractability with albumin. The penetration into the inner membrane compartment did not reach equilibrium even after 4 h at 37 degrees C. Slow penetration of a fluorescent probe such as this is useful in studies on the physico-chemical properties of the outer layer and cytoplasmic side of the platelet membranes and their changes. Initial rapid binding of the cationic probe to platelets, representing the binding of the probe to the outer layer of the plasma membrane, was increased by ionomycin-induced platelet activation. Fluorescence spectra in the presence of a relatively high concentration of the cationic probe showed increase of the excimer of the cationic probe accompanied with the incorporation of the probe to the cytoplasmic side. On ionomycin-induced activation, the excimer-to-monomer intensity ratio of the probe in the cytoplasmic side of the platelet membranes decreased, possibly due to decrease in fluidity of the lipid layer near the probe or change in distribution of the probe.
Kitagawa
S
S
Niigata College of Pharmacy, Japan.
Takegaki
M
M
D000818:Animals; D001792:Blood Platelets / Q000502:physiology*; D002417:Cattle; D005453:Fluorescence; D066298:In Vitro Techniques; D015759:Ionomycin / Q000494:pharmacology; D008051:Lipid Bilayers; D008563:Membrane Lipids / Q000502:physiology*; D015539:Platelet Activation / Q000187:drug effects; D011721:Pyrenes / Q000032:analysis / Q000378:metabolism*; D000644:Quaternary Ammonium Compounds / Q000032:analysis / Q000378:metabolism*
D008051:Lipid Bilayers; D008563:Membrane Lipids; D011721:Pyrenes; D000644:Quaternary Ammonium Compounds; D015759:Ionomycin; C076172:1-pyrenebutyltrimethylammonium