Ligand binding and expression of the main surface glycoproteins in normal mouse platelets treated with Nk protease or OSE
| mAb or ligand . | Nk treatment (% ± SEM) . | OSE treatment (% ± SEM) . |
|---|---|---|
| GPIbα (XiaG5–45kDa N-ter) | 0.01 ± 0.01 (n = 6) | 0.6 ± 0.2 (n = 9) |
| GPIbβ (RAM.1) | 100.8 (n = 2) | 97.4 ± 2.7 (n = 3) |
| GPIX (XiaB4) | 99.7 (n = 2) | 100.9 (n = 2) |
| GPV (GonC2) | 110.2 ± 11.2 (n = 3) | 95.4 (n = 2) |
| Integrin αIIbβ3 (RAM.2) | 104.9 ± 2.3 (n = 3) | 100.9 (n = 2) |
| Integrin α2β1 (SAMC1) | 109.5 ± 6.3 (n = 3) | 101.4 ± 3.9 (n = 8) |
| GPVI (JAQ1) | 101.4 ± 2.7 (n = 5) | 60.0 ± 2.9 (n = 9) |
| VWF | 0.1 (n = 2) | 0 (n = 2) |
| Thrombin (biot PPACK) | No binding in CTRL | No binding in CTRL |
| mAb or ligand . | Nk treatment (% ± SEM) . | OSE treatment (% ± SEM) . |
|---|---|---|
| GPIbα (XiaG5–45kDa N-ter) | 0.01 ± 0.01 (n = 6) | 0.6 ± 0.2 (n = 9) |
| GPIbβ (RAM.1) | 100.8 (n = 2) | 97.4 ± 2.7 (n = 3) |
| GPIX (XiaB4) | 99.7 (n = 2) | 100.9 (n = 2) |
| GPV (GonC2) | 110.2 ± 11.2 (n = 3) | 95.4 (n = 2) |
| Integrin αIIbβ3 (RAM.2) | 104.9 ± 2.3 (n = 3) | 100.9 (n = 2) |
| Integrin α2β1 (SAMC1) | 109.5 ± 6.3 (n = 3) | 101.4 ± 3.9 (n = 8) |
| GPVI (JAQ1) | 101.4 ± 2.7 (n = 5) | 60.0 ± 2.9 (n = 9) |
| VWF | 0.1 (n = 2) | 0 (n = 2) |
| Thrombin (biot PPACK) | No binding in CTRL | No binding in CTRL |
Washed platelets from wild-type mice were treated for 30 minutes with Nk protease (10 μg/mL) or OSE (48 μg/mL) and analyzed by flow cytometry for binding of mAbs against specific glycoproteins or binding of GPIbα ligands (VWF, thrombin). Results are from separate experiments as indicated and are expressed as mean percentage values (± SEM) relative to the binding in control (CTRL) untreated platelets after correction for nonspecific binding.
GP indicates glycoprotein; mAb, monoclonal antibody; OSE, O-sialoglycoprotein endopeptidase; PPACK, D-phenylalanyl-L-prolyl-L-arginine chloromethyl ketone; and VWF, von Willebrand factor.