|Title:||Analysis of human platelet glycoprotein IIb-IIIa by fluorescein isothiocyanate-conjugated disintegrins flow cytometry||Authors:||Liu C.-Z.
|Issue Date:||1994||Journal Volume:||72||Journal Issue:||6||Start page/Pages:||919-925||Source:||Thrombosis and Haemostasis||Abstract:||
Disintegrins are a group of snake venom peptides which inhibit human platelet aggregation by acting as glycoprotein IIb-IIIa (GPIIb-IIIa) antagonists. They are cysteine-rich, Arg-Gly-Asp(RGD)-containing peptides, and bind to GPIIb-IIIa complex on platelet membrane with a very high affinity (Kd, 10-7 ~ 10-8 M). In this study, we analyzed GPIIb-IIIa complex on platelet membrane by flow cytometry using fluorescein isothiocyanate (FITC)-conjugated disintegrins as probes. Of these FITC-conjugated disintegrins, FITC-Rhodostomin is the most sensitive probe because Rhodostomin was conjugated with more FITC molecules than Trigramin and Halysin were. The binding fluorescence intensity of FITC-Trigramin (FITC-Tg), FITC-Halysin (FITC-Hy) and FITC-Rhodostomin (FITC-Rn) was measured in both resting and ADP-activated platelets of diluted human platelet-rich plasma. The binding fluorescence of FITC-disintegrins was abolished by EDTA and 7E3, a monoclonal antibody against GPIIb-IIIa. ADP markedly increased the fluorescence intensity of FITC-Tg and FITC-Hy bound on platelets especially when lower doses of these probes were used, whereas it had little effect on that of FITC-Rn. Therefore, FITC-Tg and FITC-Hy can be used for the detection of the activated platelets as noted by a higher ratio of fluorescence intensity (approx. 2-4) between ADP-activated and resting platelets as compared with that (approx. 1-1.3) in the case of FITC-Rn as the probe. The platelets from three patients with Glanzmann's thrombasthenia were probed with FITC-disintegrins. As a result these three patients could be classified as type I thrombasthenia based on the extremely low level of GPIIb-IIIa detected by this method ( < 5% of normal value).
|URI:||https://scholars.lib.ntu.edu.tw/handle/123456789/564221||ISSN:||3406245||DOI:||10.1055/s-0038-1648984||SDG/Keyword:||adenosine diphosphate; edetic acid; fibrinogen receptor; fluorescein isothiocyanate; monoclonal antibody; peptide; snake venom; trigramin; article; binding affinity; case report; controlled study; disease classification; flow cytometry; fluorescence; Glanzmann disease; human; human cell; priority journal; protein binding; thrombocyte activation; thrombocyte aggregation; thrombocyte membrane
|Appears in Collections:||藥理學科所|
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