Affinity of various naturally occurring Bn-related peptides, synthetic GRP receptor (GRP-R) agonists and synthetic Bn receptor (BN-R) antagonists for NCI-N417 cells or cells transfected with hBRS-3
| Peptide No. | Peptides Added | Ki (nM) | ||
|---|---|---|---|---|
| NCI-N417 | hBRS-3-Transfected Cellsa | |||
| BALB 3T3 | NCI-H1299 | |||
| Natural Agonist Peptides | ||||
| 1 | Bn | >10,000 | >10,000 | >10,000 |
| 2 | GRP | >10,000 | >10,000 | >10,000 |
| 3 | NMB | >10,000 | 4800 ± 400 | 2800 ± 900 |
| 4 | Litorin | >10,000 | 4100 ± 400 | 3900 ± 500 |
| 5 | Phyllolitorin | >10,000 | 2100 ± 200 | 3600 ± 1300 |
| 6 | Rohdei-litorin | >10,000 | 3100 ± 700 | 2700 ± 300 |
| 7 | SAP-Bn | >10,000 | 7100 ± 950 | >10,000 |
| 8 | [Phe13]Bn | >10,000 | 6600 ± 1750 | 7100 ± 1250 |
| 9 | Frog GRP-10 | >10,000 | >10,000 | 8800 ± 300 |
| 10 | Ranatensin | 8600 ± 2000 | 6900 ± 2000 | 7400 ± 1400 |
| 11 | Alytesin | >10,000 | 3600 ± 1200 | 5100 ± 1400 |
| 12 | XenopusNMB | >10,000 | 7100 ± 2500 | >10,000 |
| 13 | Neuromedin C | >10,000 | >10,000 | >10,000 |
| 14 | [Leu8]Phyllolitorin | >10,000 | 5400 ± 1900 | 6200 ± 2000 |
| Synthetic GRP-R or NMB-R Agonists | ||||
| 15 | [dPhe6,βAla11,Phe13,Nle14]Bn(6-14) | 7.4 ± 2 | 8.9 ± 1 | 4.2 ± 1 |
| 16 | [dPhe6]Bn(6-14) | >10,000 | >10,000 | 3100 ± 1004 |
| 17 | [dPhe6,dAla11,Leu14]Bn(6-14) | >10,000 | >10,000 | 1400 ± 200 |
| 18 | [dPhe1,Nle9]litorin | 5700 ± 500 | 3200 ± 300 | 2300 ± 300 |
| 19 | Ac-NMB(3-10) | 4200 ± 400 | 820 ± 200 | 3600 ± 400 |
| Synthetic Bn-R Antagonists | ||||
| 20 | [dPhe12]Bn | 7600 ± 1000 | >10,000 | 1700 ± 400 |
| 21 | [Leu13,ψ(13-14),Leu14]Bn | >10,000 | >10,000 | 7600 ± 2100 |
| 22 | [dPhe6,Leu13,ψ(13-14),Cap14]Bn | 5000 ± 500 | >10,000 | 1100 ± 200 |
| 23 | [(3-Ph-Pr6)-His7,dAla11,dPro13,ψ(13-14),Phe14]Bn(6-14)NH2 | 4800 ± 1500 | 6800 ± 900 | 2300 ± 500 |
| 24 | [(3-Ph-Pr6)-Pro7,dAla11,dPro13,ψ(13-14),Phe14]Bn(6-14) | 4300 ± 80 | 7800 ± 700 | 3000 ± 400 |
| 25 | [dPhe6]Bn(1-13)NH2 | >10,000 | >10,000 | 2400 ± 500 |
| 26 | [dPhe6]Bn(6-13)NH2 | >10,000 | >10,000 | 5700 ± 2400 |
| 27 | [dPhe6]Bn(6-13)methyl ester | >10,000 | >10,000 | 2800 ± 900 |
| 28 | [dPhe6,dAla11]Bn(6-13)methyl ester | >10,000 | 5300 ± 2000 | 2800 ± 700 |
| 29 | [dPhe6,n-Me-dAla11]Bn(6-13)methyl ester | >10,000 | >10,000 | 4500 ± 800 |
| 30 | [dPhe6]Bn(6-13)propylamide | 5200 ± 40 | 1900 ± 300 | 2200 ± 500 |
| 31 | [dPhe6,Phe13]Bn(6-13)propylamide | >10,000 | 4500 ± 700 | 3000 ± 600 |
| 32 | [dPhe6]Bn(6-13)hexylamide | >10,000 | 3200 ± 400 | 1700 ± 600 |
| 33 | [dTyr6,dAla11]Bn(6-13)butylamide | >10,000 | 5300 ± 1400 | 2400 ± 800 |
| 34 | [dPhe1,βLeu8,des Met9]litorin | >10,000 | >10,000 | >10,000 |
| 35 | [dArg1,dTrp7,9,Leu11]Substance P | >10,000 | >10,000 | >10,000 |
| 36 | [dPro4,dTrp7,9,10]Substance P(4-11) | 3600 ± 400 | 2300 ± 400 | 1130 ± 90 |
| 37 | [dNal,Cys,Tyr,dTrp,Lys,Val,Cys,NalNH2] | 1000 ± 80 | 2800 ± 200 | 2200 ± 500 |
Cells (1–10 × 106/ml) were incubated with 75 pM125I-[dTyr6,βAla11,Phe13,Nle14]Bn(6-14) for 45 min at 22°C. Increasing concentrations of unlabeled peptide were added, and the dose-inhibition curves were analyzed by a least-squares curve-fitting program (LIGAND). Kivalues were calculated using the method of Cheng and Prusoff (Cheng and Prusoff, 1973) and are the means ± S.E. from at least three experiments performed in duplicate. a Binding data for hBRS-3 transfected cells are from (Mantey et al., 1997); >10,000 means the affinity was greater than 10,000 nM.