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Sedimentation velocity analysis of oligomeric enzymes in hydrated reversed micelles of surfactants in organic solvents

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Book cover Analytical Ultracentrifugation V

Part of the book series: Progress in Colloid and Polymer Science ((PROGCOLLOID,volume 113))

Abstract

The oligomeric state and formation of supramolecular structures of glycogen phosphorylase b from rabbit skeletal muscles have been studied in the system of hydrated reversed micelles of sodium bis-2-ethylhexyl sulfosuccinate (aerosol OT, AOT) in octane. Sedimentation analysis shows that the oligomeric state of the enzyme is controlled by the degree of hydration of the micelles ([H2O]/ [AOT] = w 0). The monomeric (in the range of w 0 from 10 to 16), dimeric (10 < w 0 < 30), trimeric (30 < w 0 < 38), tetrameric (23 < w 0 < 42), hexameric (41 < w 0 < 50), or octameric forms (48 < w 0 < 53) of the enzyme were observed depending on the degree of hydration. Sedimentation behaviour of uridine phosphorylase from Esc-herichia coli K-12 in the micellar system was studied in the range of w 0 from 8.4 to 23.9. The monomeric (at w 0 = 8.4), dimeric (at w 0 = 12.9), trimeric (at w 0 = 16.1), tetrameric (at w 0 = 18.6) and hexameric (at w 0 = 23.9) enzyme forms were registered. The results obtained show that the hydrated reversed micelles are a powerful tool for the study of not only dissociated forms of oligomeric enzymes but also supra-molecular structures. These latter structures mimic the ordered supramolecular complexes of the enzymes whose formation is favoured by the crowded molecular conditions encountered in vivo.

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Authors and Affiliations

  1. A.N. Bach Institute of Biochemistry, Russian Academy of Sciences, Leninsky prospekt 33, Moscow, 117071, Russia

    N. A. Chebotareva, B. I. Kurganov & A. A. Burlakova

Authors
  1. N. A. Chebotareva
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  2. B. I. Kurganov
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  3. A. A. Burlakova
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Correspondence to N. A. Chebotareva .

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Helmut Cölfen

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© 1999 Springer-Verlag

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Chebotareva, N.A., Kurganov, B.I., Burlakova, A.A. (1999). Sedimentation velocity analysis of oligomeric enzymes in hydrated reversed micelles of surfactants in organic solvents. In: Cölfen, H. (eds) Analytical Ultracentrifugation V. Progress in Colloid and Polymer Science, vol 113. Springer, Berlin, Heidelberg. https://doi.org/10.1007/3-540-48703-4_18

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  • DOI: https://doi.org/10.1007/3-540-48703-4_18

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  • Publisher Name: Springer, Berlin, Heidelberg

  • Print ISBN: 978-3-540-66175-7

  • Online ISBN: 978-3-540-48703-6

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