Physicochemical characterization of cross-linked human serum albumin dimer and its synthetic heme hybrid as an oxygen carrier

Teruyuki Komatsu, Yukiko Oguro, Yuji Teramura, Shinji Takeoka, Junpei Okai, Makoto Anraku, Masaki Otagiri, Eishun Tsuchida

    Research output: Contribution to journalArticle

    32 Citations (Scopus)

    Abstract

    The recombinant human serum albumin (rHSA) dimer, which was cross-linked by a thiol group of Cys-34 with 1,6-bis(maleimido)hexane, has been physicochemically characterized. Reduction of the inert mixed-disulfide of Cys-34 beforehand improved the efficiency of the cross-linking reaction. The purified dimer showed a double mass and absorption coefficient, but unaltered molar ellipticity, isoelectric point (pI: 4.8) and denaturing temperature (65°C). The concentration dependence of the colloid osmotic pressure (COP) demonstrated that the 8.5 g dL -1 dimer solution has the same COP with the physiological 5 g dL -1 rHSA. The antigenic epitopes of the albumin units are preserved after bridging the Cys-34, and the circulation lifetime of the 125I-labeled variant in rat was 18 h. A total of 16 molecules of the tetrakis{(1-methylcyclohexanamido)phenyl}porphinatoiron(II) derivative (FecycP) is incorporated into the hydrophobic cavities of the HSA dimer, giving an albumin-heme hybrid in dimeric form. It can reversibly bind and release O 2 under physiological conditions (37°C, pH 7.3) like hemoglobin or myoglobin. Magnetic circular dichroism (CD) revealed the formation of an O 2-adduct complex and laser flash photolysis experiments showed the three-component kinetics of the O 2-recombination reaction. The O 2-binding affinity and the O 2-association and -dissociation rate constants of this synthetic hemoprotein have also been evaluated.

    Original languageEnglish
    Pages (from-to)21-31
    Number of pages11
    JournalBiochimica et Biophysica Acta - General Subjects
    Volume1675
    Issue number1-3
    DOIs
    Publication statusPublished - 2004 Nov 18

    Fingerprint

    Osmotic Pressure
    Colloids
    Heme
    Serum Albumin
    Dimers
    Albumins
    Oxygen
    Myoglobin
    Photolysis
    Cross Reactions
    Isoelectric Point
    Hexanes
    Circular Dichroism
    Sulfhydryl Compounds
    Disulfides
    Genetic Recombination
    Epitopes
    Hemoglobins
    Lasers
    Temperature

    Keywords

    • Albumin-heme dimer
    • Colloid osmotic pressure
    • Cross-linking
    • Human serum albumin dimer
    • Oxygen carrier
    • Synthetic heme

    ASJC Scopus subject areas

    • Biochemistry
    • Biophysics
    • Molecular Biology

    Cite this

    Physicochemical characterization of cross-linked human serum albumin dimer and its synthetic heme hybrid as an oxygen carrier. / Komatsu, Teruyuki; Oguro, Yukiko; Teramura, Yuji; Takeoka, Shinji; Okai, Junpei; Anraku, Makoto; Otagiri, Masaki; Tsuchida, Eishun.

    In: Biochimica et Biophysica Acta - General Subjects, Vol. 1675, No. 1-3, 18.11.2004, p. 21-31.

    Research output: Contribution to journalArticle

    Komatsu, Teruyuki ; Oguro, Yukiko ; Teramura, Yuji ; Takeoka, Shinji ; Okai, Junpei ; Anraku, Makoto ; Otagiri, Masaki ; Tsuchida, Eishun. / Physicochemical characterization of cross-linked human serum albumin dimer and its synthetic heme hybrid as an oxygen carrier. In: Biochimica et Biophysica Acta - General Subjects. 2004 ; Vol. 1675, No. 1-3. pp. 21-31.
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    AU - Oguro, Yukiko

    AU - Teramura, Yuji

    AU - Takeoka, Shinji

    AU - Okai, Junpei

    AU - Anraku, Makoto

    AU - Otagiri, Masaki

    AU - Tsuchida, Eishun

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