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Reversible modification of thiol-containing polypeptides with poly(ethylene glycol) through formation of mixed disulfide bonds

The case of papaya proteinase III

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Abstract

Papaya proteinase III (PPIII) was purified, as the S-methylthio derivative from the latex ofCarica papaya L., by ion-exchange chromatography. Separation of reactivable PPIII from the irreversibly oxidized molecular species of this enzyme was readily achieved after a selective conversion of the reactivated proteinase into the S-monomethoxypoly-(ethylene glycol) thio derivative (S-mPEG thio PPIII). From this derivative, a PPIII preparation titrating 1 mol of thiol/mol of enzyme was regenerated. From the physicochemical properties of S-mPEG thio PPIII that were investigated, it is concluded that interactions between the mPEG and the PPIII chains occur only to a limited extent. In addition to its usefulness for purifying thiol-containing enzymes, the mPEG modification resulting from mixed disulfide bond formation may find other practical applications.

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Authors and Affiliations

  1. Protein Chemistry Unit, Service de Chimie générale 1, Faculté de Médecine, Université Libre de Bruxelles, Campus de la Plaine, Boulevard du Triomphe, B-1050, Bruxelles, Belgium

    Tony Musu, Mohamed Azarkan, Jeanne Brygier, Claudine Paul, Jean Vincentelli, Danielle Baeyens-Volant, Claude Guermant, Michelle Nijs & Yvan Looze

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  1. Tony Musu
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  2. Mohamed Azarkan
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  3. Jeanne Brygier
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  4. Claudine Paul
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  6. Danielle Baeyens-Volant
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  8. Michelle Nijs
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  9. Yvan Looze
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Musu, T., Azarkan, M., Brygier, J. et al. Reversible modification of thiol-containing polypeptides with poly(ethylene glycol) through formation of mixed disulfide bonds. Appl Biochem Biotechnol 56, 243–263 (1996). https://doi.org/10.1007/BF02786956

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  • DOI: https://doi.org/10.1007/BF02786956

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