Structure/activity relationships for TRAPs
| Peptide | Cell/Tissue Type | Effect (EC50/IC50) | References |
|---|---|---|---|
| SFLLRNPNDKYEPF (TRAP-14) | Xenopusoocytes, platelets, rat aortic rings (endothelium denuded or intact), guinea pig gastric longitudinal smooth muscle, CCL39 hamster fibroblasts, rat glomerular mesangial cells, rat astrocytes | 4–30 μM | Chao et al., 1992; Coller et al., 1992; Kawabata et al., 1999c;Vouret-Craviari et al., 1992; Vu et al., 1991a; Yang et al., 1992 |
| SFLLR-NH2 | Platelets, transfected mammalian cells, endothelium denuded and intact RA, CCL39 fibroblasts | 0.5–6 μM | Ceruso et al., 1999; Hollenberg et al., 1996; Kawabata et al., 1999c; Laniyonu and Hollenberg, 1995; Natarajan et al., 1995;Scarborough et al., 1992 |
| NPNDKYEPF short peptides (<5) | Platelets CCL39 fibroblasts, various species platelets, rat glomerular mesangial cells, endothelium denuded RA and gastric LM | >200 μM Loss of function |
Vassallo et al., 1992
Albrightson et al., 1994; Bernatowicz et al., 1996; Chao et al., 1992; Connolly et al., 1994; Vouret-Craviari et al., 1992 |
| Substituted peptides | |||
| Acetyl-SFLLR-NH2 | Platelets, transfected mammalian cells, rat gomerular mesengial, SH-EP cells | >1000 μM | Albrightson et al., 1994; Coller et al., 1992; Sakaguchi et al., 1994; Scarborough et al., 1992 |
| H-SFLLR-NH2 | Platelets, transfected mammalian cells, SH-EP cells | Low activity | Scarborough et al., 1992; Shimohigashi et al., 1994; Van Obberghen-Schilling et al., 1993 |
| XFFLR-NH2 | Various cell types | Charged amino acids not tolerated, size/shape important; Thr substitution yields a PAR-1-specific peptide | Bischoff et al., 1994;Ceruso et al., 1999; Chao et al., 1992; Hollenberg et al., 1997;Natarajan et al., 1995; Sakaguchi et al., 1994; Scarborough et al., 1992; Van Obberghen-Schilling et al., 1993; Vassallo et al., 1992; Yang et al., 1992 |
| SXLLR-NH2 | Various cell types | Only aromatic residues tolerated | Albrightson et al., 1994;Ceruso et al., 1999; Chao et al., 1992; Natarajan et al., 1995; Nose et al., 1993; Scarborough et al., 1992; Van Obberghen-Schilling and Pouyssegur, 1993; Vassallo et al., 1992 |
| SFXLR-NH2 | Various cell types | No loss of activity, more active with 3-(2-naphthyl)-l-alanine | Bischoff et al., 1994;Blackhart et al., 1996; Ceruso et al., 1999; Chao et al., 1992;Laniyonu and Hollenberg, 1995; Natarajan et al., 1995; Scarborough et al., 1992; Shimohigashi et al., 1994; Van Obberghen-Schilling and Pouyssegur, 1993; Vassallo et al., 1992 |
| SFLXR-NH2 | Various cell types | Acidic and basic amino acids not tolerated | Blackhart et al., 1996; Ceruso et al., 1999; Chao et al., 1992; Natarajan et al., 1995; Scarborough et al., 1992; Vassallo et al., 1992 |
| SFLLX-NH2 | Various cell types | Wide range of residues tolerated, but reduced activity | Blackhart et al., 1996; Ceruso et al., 1999; Chao et al., 1992; Hollenberg et al., 1997;Natarajan et al., 1995; Nose et al., 1998b; Scarborough et al., 1992;Vassallo et al., 1992 |
| Monocyclic SFLLRN analogues | Platelets | Significant loss of activity | McComsey et al., 1999 |
| Improved agonists | |||
| S(p-F)FFLRNP | Platelets, SH-EP cells | 1.7 μM | Nose et al., 1998a; Nose et al., 1993; Shimohigashi et al., 1994 |
| S(p-F)FpGuFLR-NH2 | Platelets, smooth muscle, mouse fibroblasts | 0.4 μM | Bernatowicz et al., 1996 |
| A(p-F)FRCha-HarY-NH2 | Platelets | 0.01–0.12 μM | Ahn et al., 1997; Debeir et al., 1997; Feng et al., 1995;Kawabata et al., 1999c |
| A(p-F)FRChaCitY-NH2 | Platelets | 0.2 μM | Kawabata et al., 1999c |
| S(p-F)FHarLRK-NH2 | Xenopusoocytes | 0.05–0.1 μM | Blackhart et al., 1996 |
| S(p-F)F2NaALR-NH2 | Platelets, CHRF-288 membranes | 80 nM | Seiler et al., 1996 |
| Antagonists | IC50 | ||
| 1-phenylacetyl-4-(6-guanidohexanoyl)-piperazine | Platelets | 50% inhibition of SFLLRNP | Alexopoulos et al., 1998 |
| 1-(6-guanidohexanoyl)- 4-(phenylacetylamido-methyl)-piperazidine | Platelets | 40% inhibition of SFLLRN | Alexopoulos et al., 1998 |
| BMS-197525 {N-trans(p-F)FpGuFLR-NH2} | Platelets, smooth muscle, mouse fibroblasts | 0.2 μM of SFLLRNP | Bernatowicz et al., 1996 |
| BMS-200261 {N-trans(p-F)FpGuFLRR-NH2} | Platelets | 20 nM (SFLLRN) 1.6 μM (thrombin) | Bernatowicz et al., 1996; Kawabata et al., 1999c |
| 3-Mercapto-propionyl-FChaChaRKNDK-NH2 | Platelets | 0.7–6.4 μM (thrombin) | Kawabata et al., 1999c; Seiler et al., 1995 |
| LVR(D-)CGKHSR | Rat astrocytes | 180 μM ([3H]thymidine incorporation, by TRAP14 or thrombin) | Debeir et al., 1997 |
| Oxazole-30 | Platelets, CHRF membranes | 25 μM (thrombin), 6.6 μM (SFLLRN) | Hoekstra et al., 1998 |
| S(Npys)-βMp-(p-F)F-NHCH(C6H5)2 | Platelets | 52 μM (SFLLRNP) | Fujita et al., 1999 |
| S(Npys)-βMp-(p-F)F-NHCH2CH(C6H5)2 | Platelets | 54 μM (SFLLRNP) | Fujita et al., 1999 |
| RWJ-56110 series | Platelets | 0.34 μM (thrombin) 0.16 μM (SFLLRN) | Andrade-Gordon et al., 1999 |
| SCH 79797 | Platelets, smooth muscle | 70 nM ([3H]haTRAP binding) | Ahn et al., 2000 |
| SCH 203099 | Platelets, smooth muscle | 45 nM ([3H]haTRAP binding) | Ahn et al., 2000 |
| FR171113 | Platelets | 0.29 μM (thrombin) | Kato et al., 1999 |
| Labels | IC50/K d | ||
| SFp-azido-FLRNPKGGK-biotin | HEL cells | No effect | Bischoff et al., 1994 |
| [3H]haTRAP {[3H]A(p-F)FRChaHarY-NH2} | Platelet membranes, platelets | 0.15 μM, K d = 15 nM | Ahn et al., 1997 |
| A(p-F)FRChaHar(125I)Y-NH2 | Platelets | 0.03 μM | Feng et al., 1995 |
| BMS-200661 {N-trans(p-F)FpGuFLROrn} | Platelets | K d= 10–30 nM | Bernatowicz et al., 1996 |
| SFLLRNPNDKYEPF-biotin | BHK cells | K d = 3 μmol/l | Takada et al., 1995 |
| BMS-197525, [3H] and biotinylated derivatives | CHRF-288 cells | K d= 80 nM or less | Elliott et al., 1999 |
Amino acid residues X indicates amino acid scan; Har, homoarginine; (pF)F, parafluorophenylalanine; Cha, cyclohexylalanine;N-trans, trans-cinnomoyl; pGuF,p-guanidino-phenylalanine; Cit, citrulline; Orn, ornithine; 2NaA, 2-naphthylalanine; Npys,S-3-nitro-2-pyridinesulphenyl; β-Mp, β-mercaptopropionyl; [3H]haTRAP, [3H]A(p-F)FRChaHarY-NH2. All EC50values refer to platelet aggregation unless indicated otherwise. Likewise, all IC50 values refer to inhibition of TRAP-induced platelet aggregation unless indicated otherwise.