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On the interaction of rabbit antithrombin III with the luminal surface of
the normal and deendothelialized rabbit thoracic aorta in vitro
MW Hatton, SL Moar and M Richardson
Pure rabbit antithrombin III was isotope labeled (with 125I or 3H) by two
different methods; neither procedure caused a loss of antithrombin activity
although both methods affected the affinity of the protein for
Sepharose-heparin. From segments from freshly excised rabbit aorta, the
uptake of isotope-labeled antithrombin III by the endothelium was rapid and
saturable, although relatively small compared to the uptake of thrombin;
binding of 3H-antithrombin III to the endothelium resembled that of
125I-antithrombin III. Transendothelial passage of antithrombin III into
the subendothelial layers (intima-media) was slow and progressive.
Endothelium binding was not affected by pretreating the vessel with either
heparin, thrombin, or glycosaminoglycan-specific enzymes. Endothelium-bound
antithrombin III was not selectively displaced by either heparin or
thrombin. In contrast, endothelium-bound thrombin was rapidly dislodged by
antithrombin III as a thrombin- antithrombin III complex. The surface of
the deendothelialized aorta (ie, subjected to a balloon catheter) bound
antithrombin III avidly. Pretreatment of the deendothelialized vessel with
glycosaminoglycan- specific enzymes, particularly heparitinase, decreased
intima-media binding by up to 80%. 125I-antithrombin III, when bound to the
deendothelialized vessel surface, was actively displaced by either heparin,
thrombin, or by unlabeled antithrombin III. The relatively poor binding of
antithrombin III compared with that of thrombin by the endothelium in vitro
supports an earlier proposal (Lollar P, Owen WG: J Clin Invest
66:1222-1230, 1980) that thrombin bound to high-affinity sites, possibly
pericellular proteoglycan, of the endothelium is inactivated by plasma
antithrombin III in vivo. Such a situation probably holds for large
arteries at least.
Volume 67,
Issue 4,
pp. 878-886,
04/01/1986
Copyright © 1986 by The American Society of Hematology
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