Abstract
A method for the modification of enzymes by MPEG carrying an amino acid or peptide as a spacer arm is described and tested with aliphatic or aromatic side chains amino acids. The procedure involves MPEG activation byp-nitrophenylchloroformate for the amino acid or peptide coupling that is in turn activated for the protein binding.
The advantage of the method resides in the possibility to introduce proper reporter groups between the polymer and the protein as norleucine for a direct evaluation of the bound polymer chains, tryptophan for structural studies of the polymer-protein adduct, and radioactive amino acid for pharmacokinetic investigations.
The method was positively tested with arginase, ribonuclease, and superoxide dismutase as enzymes of therapeutic value.
Similar content being viewed by others
References
Abuchowski, A. and Davis, F. F. (1981), inEnzymes as Drugs, Holcenberg, J. S. and Roberts, J., ed., Wiley & Sons, pp 367-384.
Conforti, A., Franco, L., Milanino, R., Velco, G. P., Boccu, E., Largajolli, R., Schiavon, O. and Veronese, F. M. (1987),Pharmacol. Research Comunic. 19, 287.
Veronese, F. M. (1989),Chimica Oggi 7, 53.
Yoshinga, K. and Harris, J. M. (1989),J. Bioactive Compatible Polymers 4, 17.
Kopecek, J. and Duncan, R. (1987),J. Controlled Release 6, 315.
Crook, E. M., Mathias, A. P. and Rabin, B. R. (1960),J. Biochem. 74, 234.
Paoletti, F., Aldinucci, D., Mocali, A. and Caparrini, A. (1986),Anal. Biochem. 154, 536.
Snyder, S. L. and Sabocinsky, P. Z. (1975),Anal. Biochem. 64, 248.
Sims, G. E. C. and Snape, T. J. (1980),Anal. Biochem. 107, 60.
Visco, C, Benassi, C. A., Veronese, F. M., and Miglioli, P. A. (1987),II Farmaco 42, 549.
Caliceti, P., Schiavon, O., Mocali, A., and Veronese, F. M. (1989),II Farmaco 44, 711.
Veronese, F.M., Caliceti, P., Pastorino, A., Schiavon, O., Sartore, L., Banci, L., and Monsu Scolaro, L. (1989),J. Controlled Release 10, 145.
Schiavon, O. et al. in preparastion.
Joppich, M., and Luisi, P. L. (1979),Makromol. Chem. 180, 1381.
Brand, L. and Whithold, B. (1967),Methods Enzymol. 11, 776.
Veronese, F. M., Sartore, L., Caliceti, P., Schiavon, O., Ranucci, E., and Ferruti, P. (1989),J. Bioactive Compatible Polymers (in press).
Hemant, N. J. (1988),Pharmaceutical Technology 12, 118.
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Sartore, L., Caliceti, P., Schiavon, O. et al. Enzyme modification by MPEG with an amino acid or peptide as spacer arms. Appl Biochem Biotechnol 27, 45–54 (1991). https://doi.org/10.1007/BF02921514
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF02921514