A Role for Pericellular Proteoglycan in the Binding of Thrombin or Antithrombin III by the Blood Vessel Endothelium? – The Effects of Proteoglycan-Degrading Enzymes and Glycosaminoglycan-Binding Proteins on ¹²⁵I-Thrombin Binding by the Rabbit Thoracic Aorta In Vitro

Summary

Rabbit thoracic aorta segments were treated with either proteoglycan-degrading enzymes or with glycosaminoglycan- binding proteins to examine the nature of the endothelial and subendothelial binding sites of ¹²⁵I-thrombin. Treatment (5-30 min) with enzymes (heparitinase, chondroitinases AC or ABC) caused a decrease in ¹²⁵I-thrombin binding by the endothelium (30-70%) and by the subendothelial (intima-media) layer (20-50%); a low- specificity protease destroyed endothelial binding almost entirely and reduced binding to the subendothelium by approximately 60% over a similar period. Of the glycosaminoglycan-binding proteins, pretreatment of the aorta wall with protamine caused a 30% decrease in thrombin binding to the endothelium whereas lipoprotein lipase (present during ¹²⁵I-thrombin uptake) decreased binding by up to 40%. Pretreatment with antithrombin III did not significantly affect binding of either ¹²⁵I-thrombin or ¹²⁵I-FPR-inactivated thrombin. In contrast to thrombin, ¹²⁵I- antithrombin III was not readily uptaken by the aorta segments. These observations indicate that, whereas the minimal binding by ¹²⁵I-antithrombin III probably does not involve endothelial proteoglycan, a strong case can be made for endothelial and subendothelial proteoglycan binding sites for thrombin..

Keywords

Thrombin - Antithrombin III - Aorta - Endothelium - Intimamedia - Proteoglycan - Glycosaminoglycan

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