Arg-Tyr-Asp (RYD) and Arg-Cys-Asp (RCD) motifs in dendroaspin promote selective inhibition of beta1 and beta3 integrins.
Arg-Gly-Asp (RGD) is a unique minimal integrin-binding sequence that is found within several glycoprotein ligands. This sequence has also been found in snake-venom anti-platelet proteins, including the disintegrins and dendroaspin, a natural variant of short-chain neurotoxins isolated from the venom of Dendroaspis jamesonii. In the present study, the motifs RYD and RCD were introduced into the dendroaspin scaffold to replace RGD. Both motifs in dendroaspin caused inhibition of ADP-induced platelet aggregation with IC(50) values of 200 and 300 nM respectively, similar to that of the wild-type RGD motif (170 nM). In comparison with wild-type dendroaspin, both RYD- and RCD-containing dendroaspins were more selective in the inhibition of the adhesion of K562 cells to laminin rather than to fibrinogen and fibronectin, even though they were 10-30-fold less potent at inhibiting K562 cell (containing alpha(5)beta(1) integrin) adhesion to laminin compared with wild-type. Interestingly, the RYD motif produced a similar IC(50) value to the RGD motif at inhibiting A375-SM cell (beta(3) integrin) adhesion to collagen, whereas the RCD motif was approx. 2-6-fold less potent compared with either RGD or RYD. These findings show that the selectivity of dendroaspin binding to beta(1) and beta(3) integrins can be modulated by the introduction of alternative cell recognition sequences.
Wattam
B
B
Thrombosis Research Institute, Manresa Road, London SW3 6LR, UK.
Shang
D
D
Rahman
S
S
Egglezou
S
S
Scully
M
M
Kakkar
V
V
Lu
X
X
D020816:Amino Acid Motifs; D015703:Antigens, CD / Q000187:drug effects*; D002448:Cell Adhesion / Q000187:drug effects; D003094:Collagen; D004546:Elapid Venoms / Q000494:pharmacology*; D005340:Fibrinogen; D005353:Fibronectins; D019012:Integrin beta1 / Q000187:drug effects*; D039661:Integrin beta3; D016023:Integrins / Q000187:drug effects; D007797:Laminin; D009842:Oligopeptides; D010975:Platelet Aggregation Inhibitors / Q000494:pharmacology*; D010980:Platelet Membrane Glycoproteins / Q000187:drug effects*
D015703:Antigens, CD; D004546:Elapid Venoms; D005353:Fibronectins; D019012:Integrin beta1; D039661:Integrin beta3; D016023:Integrins; D007797:Laminin; D009842:Oligopeptides; D010975:Platelet Aggregation Inhibitors; D010980:Platelet Membrane Glycoproteins; C078264:arginyl-tyrosyl-aspartic acid; C074772:mambin; C047981:arginyl-glycyl-aspartic acid; D005340:Fibrinogen; D003094:Collagen