Table I.
Summary of CXCL8 Binding to CXCR1 N-Terminal Domain Peptides Reported in Literature
| CXCL8 variant | CXCR1 | Technique | Buffer conditions | [CXCL8], µM | Kd (µM) | Reference |
|---|---|---|---|---|---|---|
| WT dimer | h40mera | NMR | 50 mM Pi, pH 6.7 | 750 | 170 | Clubb et al., 199424 |
| WT dimer | h38mer | NMR | 20 mM HEPES, pH 5.5 | 100 | 70 | Park et al., 26,44 |
| WT dimer | h21mer | NMR | 50 mM Pi, 150 mM NaCl, pH 6.5 | ≥200 | 550 | Kendrick et al., 27 |
| WT dimerb | r24mer | NMR | 50 mM acetate, pH 6.0 | 90c | n.d. | Ravindran et al., 200923 |
| R26C dimer | r24mer | NMR | 50 mM acetate, pH 6.0 | 93c | 360 | Ravindran et al., 200923 |
| 1–66 monomer | h40mer | NMR | 50 mM acetate, pH 5.5 | 130–200 | <10 | Barter and Stone, 25 |
| 1–66 monomer | h40mer | ITC | 50 mM MOPS, pH 7.0 | 9.6 | Barter and Stone, 25 | |
| L25Y/V27R monomer | h21mer | NMR | 50 mM Pi, 150 mM NaCl, pH 6.5 | ≥200 | 460 | Kendrick et al., 27 |
| 1–66 monomer | r24mer | NMR | 50 mM acetate, pH 6.0 | 115c | 12 | Ravindran et al., 200923 |
| L25NMe monomer | r34mera | ITC | 50 mM HEPES, 50 mM NaCl, pH 8.0 | 6.0 | Fernando et al., 21 | |
| 1–66 monomer | r34mera | ITC | 50 mM Pi, 50 mM NaCl, pH 8.0 | 8.7 | Fernando et al., 22 | |
| 1–66 monomer | r34mera | ITC | 50 mM HEPES, 50 mM NaCl, pH 8.0 | 8.6 | Fernando et al., 22 | |
| 1–66 monomer | r34mera | ITC | 50 mM Tris, 50 mM NaCl, pH 8.0 | 7.5 | Fernando et al., 22 |
| CXCR1 constructs | ||||||
|---|---|---|---|---|---|---|
| h40mera | MSNITDPQMWDFDDLNFTGMPPADEDYSPSMLETETLNKY | |||||
| h38merd | MSNITDPQMWDFDDLNFTGMPPADEDYSPSMLETETLN | |||||
| h29mer | MSNITDPQMWDFDDLNFTGMPPADEDYSP | |||||
| h21mer | MWDFDDLNFTGMPPADEDYSP | |||||
| r24mer | LWTWFEDEFANATGMPPVEKDYSP | |||||
| r34mera | LWTWFEDEFANATGMPPVEKDYSPSLVVTQTLNK | |||||
| CXCL8 constructs | |
|---|---|
| WT | SAKELRCQCIKTYSKPFHPKFIKELRVIESGPHCANTEIIVKLSDGRELCLDPKENWVQRVVEKFLKRAENS |
| R26Ce | SAKELRCQCIKTYSKPFHPKFIKELCVIESGPHCANTEIIVKLSDGRELCLDPKENWVQRVVEKFLKRAENS |
| L25NMee | SAKELRCQCIKTYSKPFHPKFIKELRVIESGPHCANTEIIVKLSDGRELCLDPKENWVQRVVEKFLKRAENS |
| V27P/E29Pe | SAKELRCQCIKTYSKPFHPKFIKELRPIPSGPHCANTEIIVKLSDGRELCLDPKENWVQRVVEKFLKRAENS |
| L25Y/V27Re | SAKELRCQCIKTYSKPFHPKFIKEYRRIESGPHCANTEIIVKLSDGRELCLDPKENWVQRVVEKFLKRAENS |
| 1–66 | SAKELRCQCIKTYSKPFHPKFIKELRVIESGPHCANTEIIVKLSDGRELCLDPKENWVQRVVEKFL |
h, human; r, rabbit; Pi, phosphate.
a
native cysteine mutated to serine
b
dimer dissociation coupled to r24mer binding
c
personal communication (Ravindran, A)
d
extra GS on the N-terminus and hexa His-tag on the C-terminus
e
R26C – Arg26 is substituted with a Cys (in red); L25NMe - Leu25 (in red italic) backbone amide is substituted with a methyl group; V27P/E29P – Val27 and Glu29 are substituted with a Pro (in red); L25Y/V27R – Leu25 and Val27 are substituted with Tyr and Arg, respectively (in red).