Conjugation of von Willebrand factor-binding domain of platelet glycoprotein Ibα to size-controlled albumin microspheres

Shinji Takeoka, Yuji Teramura, Haruki Ohkawa, Yasuo Ikeda, Eishun Tsuchida

    Research output: Contribution to journalArticle

    27 Citations (Scopus)

    Abstract

    Albumin microspheres (AMS), of which the average diameter is 240 ± 10 nm, were prepared by pH control and heat treatment. Cytochrome c and rGPIbα; a water-soluble fragment of the α chain of a recombinant platelet glycoprotein (GP)Ib containing a von Willebrand factor (vWf)-binding site were selected as a receptor protein. Cytochrome c was used as a probe protein for monitoring. Onto the surface of the AMS and those proteins, N-succinimidyl 3-(2-pyridyldithio)propionate (SPDP) was reacted through the amide linkage to obtain PD-AMS, PD-cytochrome c, and PD-rGPIbα, respectively. The latter two were further reduced to SH-cytochrome c and SH-rGPIbα by dithiothreitol and conjugated with PD-AMS by a thiol - disulfide exchange reaction. The resulting AMS contain cytochrome c or rGPIbα of about 25000 or 2500 molecules, respectively. The addition of ristocetin to the rGPIbα-AMS in the presence of vWf caused specific aggregation. Furthermore, the rGPIbα-AMS enhanced the ristocetin induced platelet aggregation in a low platelet concentration (4.0 × 107/mL).

    Original languageEnglish
    Pages (from-to)290-295
    Number of pages6
    JournalBiomacromolecules
    Volume1
    Issue number2
    Publication statusPublished - 2000 Jun

    Fingerprint

    Platelet Glycoprotein GPIb-IX Complex
    Platelet Membrane Glycoproteins
    Glycoproteins
    von Willebrand Factor
    Platelets
    Microspheres
    Albumins
    Cytochromes c
    Proteins
    Ristocetin
    Agglomeration
    Dithiothreitol
    Platelet Aggregation
    Sulfhydryl Compounds
    Amides
    Disulfides
    Binding sites
    Blood Platelets
    Hot Temperature
    Binding Sites

    ASJC Scopus subject areas

    • Biochemistry, Genetics and Molecular Biology(all)
    • Organic Chemistry
    • Materials Chemistry
    • Polymers and Plastics

    Cite this

    Conjugation of von Willebrand factor-binding domain of platelet glycoprotein Ibα to size-controlled albumin microspheres. / Takeoka, Shinji; Teramura, Yuji; Ohkawa, Haruki; Ikeda, Yasuo; Tsuchida, Eishun.

    In: Biomacromolecules, Vol. 1, No. 2, 06.2000, p. 290-295.

    Research output: Contribution to journalArticle

    Takeoka, Shinji ; Teramura, Yuji ; Ohkawa, Haruki ; Ikeda, Yasuo ; Tsuchida, Eishun. / Conjugation of von Willebrand factor-binding domain of platelet glycoprotein Ibα to size-controlled albumin microspheres. In: Biomacromolecules. 2000 ; Vol. 1, No. 2. pp. 290-295.
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