Inhibition of platelet aggregation by grafting RGD and KGD sequences on the structural scaffold of small disulfide-rich proteins.

Disintegrins represent a group of disulfide-rich peptides ranging in size from 41 to over 80 residues and are antagonists of several integrin receptors. Disintegrins containing an RGD or KGD sequence are potent inhibitors of platelet aggregation as they block the binding of fibrinogen to alpha(IIb)beta(3) integrin. The high affinity binding to alpha(IIb)beta(3) in comparison to short linear peptides has been attributed to the localisation of the RGD or KGD sequence within a defined three-dimensional structure. Cystine knot microproteins are members of another family of small disulfide-rich peptides that consist of only 28-40 amino acid residues. They display numerous biological activities depending on the peptide sequence of loop regions that are fixed on a structural scaffold that is stabilised by three knot-forming disulfide bonds. In the present study we grafted RGD and KGD containing peptide sequences with seven and 11 amino acids, respectively, into two cystine knot microproteins, the trypsin inhibitor EETI-II and the melanocortin receptor binding domain of the human agouti-related protein AGRP, as well as into the small disintegrin obtustatin. The engineered proteins were much more potent to inhibit the fibrinogen binding, alpha(IIb)beta(3) activation and platelet aggregation when compared to the grafted peptides. Differences that were observed between the engineered proteins indicate the importance of the structural scaffold and the amino acids neighbouring the grafted peptide sequences.

The role of platelet receptors and adhesion molecules in coronary artery disease.

Platelets play a significant role in coronary artery disease through interactions with each other and with other cell types. These interactions are mediated by certain receptors on the surface of platelets and other cells which can lead to intra-coronary thrombus formation and occlusion that may result in acute coronary syndromes. The important roles of the currently available anti-platelet therapies have been well established in many clinical outcome trials in cardiovascular patients. An understanding of these different interactions provides the clinician with a background that supports the clinical importance of currently available anti-platelet therapies. Moreover, knowledge of the mechanisms of cellular crosstalk will lead to important advances in the development of better antithrombotic therapies.