Reversible Modification of Thiol-containing Polypeptides with Poly (ethylene Glycol) Through Formation of Mixed Disulfide Bonds. The Case of Papaya Proteinase III

Overview

Journal Appl Biochem Biotechnol

Specialties Biochemistry
Biotechnology

Date 1996 Mar 1

PMID 8984898

Citations 1

Authors

T Musu

M Azarkan

J Brygier

C Paul

J Vincentelli

C Guermant

M Nijs

Y Looze

Affiliations

Soon will be listed here.

Abstract

Papaya proteinase III (PPIII) was purified, as the S-methylthio derivative from the latex of Carica 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.

Citing Articles

Purification and autolysis of the ficin isoforms from fig (Ficus carica cv. Sabz) latex.

Zare H, Moosavi-Movahedi A, Salami M, Mirzaei M, Saboury A, Sheibani N Phytochemistry. 2013; 87:16-22.

PMID: 23312458 PMC: 3755362. DOI: 10.1016/j.phytochem.2012.12.006.

References

Walter H, RAYMOND F, Fisher D . Erythrocyte partitioning in dextran-poly(ethylene glycol) aqueous phase systems. Events in phase and cell separation. J Chromatogr. 1992; 609(1-2):219-27. DOI: 10.1016/0021-9673(92)80166-r. View

Zalipsky S, Seltzer R . Evaluation of a new reagent for covalent attachment of polyethylene glycol to proteins. Biotechnol Appl Biochem. 1992; 15(1):100-14. DOI: 10.1111/j.1470-8744.1992.tb00198.x. View

Laemmli U . Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970; 227(5259):680-5. DOI: 10.1038/227680a0. View

Abuchowski A, van Es T, Palczuk N, DAVIS F . Alteration of immunological properties of bovine serum albumin by covalent attachment of polyethylene glycol. J Biol Chem. 1977; 252(11):3578-81. View

Goormaghtigh E, Cabiaux V, Ruysschaert J . Secondary structure and dosage of soluble and membrane proteins by attenuated total reflection Fourier-transform infrared spectroscopy on hydrated films. Eur J Biochem. 1990; 193(2):409-20. DOI: 10.1111/j.1432-1033.1990.tb19354.x. View

Shipton M, Brochlehurst K . Characterization of the papain active centre by using two-protonic-state electrophiles as reactivity probes. Evidence for nucleophilic reactivity in the un-interrupted cysteine-25-histidine-159 interactive system. Biochem J. 1978; 171(2):385-401. PMC: 1183967. DOI: 10.1042/bj1710385. View

Jacquet A, Kleinschmidt T, Dubois T, SCHNEK A, Looze Y, Braunitzer G . The thiol proteinases from the latex of Carica papaya L. IV. Proteolytic specificities of chymopapain and papaya proteinase omega determined by digestion of alpha-globin chains. Biol Chem Hoppe Seyler. 1989; 370(8):819-29. DOI: 10.1515/bchm3.1989.370.2.819. View

Baker E . Structure of actinidin, after refinement at 1.7 A resolution. J Mol Biol. 1980; 141(4):441-84. DOI: 10.1016/0022-2836(80)90255-7. View

Stocks S, Jones A, Ramey C, Brooks D . A fluorometric assay of the degree of modification of protein primary amines with polyethylene glycol. Anal Biochem. 1986; 154(1):232-4. DOI: 10.1016/0003-2697(86)90520-8. View

10.

Lowe G, Whitworth A . A kinetic and fluorimetric investigation of papain modified at tryptophan-69 and -177 by N-bromosuccinimide. Biochem J. 1974; 141(2):503-15. PMC: 1168105. DOI: 10.1042/bj1410503. View

11.

Goodson R, Katre N . Site-directed pegylation of recombinant interleukin-2 at its glycosylation site. Biotechnology (N Y). 1990; 8(4):343-6. DOI: 10.1038/nbt0490-343. View

12.

Buttle D, Kembhavi A, Sharp S, Shute R, Rich D, Barrett A . Affinity purification of the novel cysteine proteinase papaya proteinase IV, and papain from papaya latex. Biochem J. 1989; 261(2):469-76. PMC: 1138849. DOI: 10.1042/bj2610469. View

13.

Skuse D, Yalpani M, Brooks D . Hydroxypropyl cellulose/poly(ethylene glycol)-co-poly(propylene glycol) aqueous two-phase systems: system characterization and partition of cells and proteins. Enzyme Microb Technol. 1992; 14(10):785-90. DOI: 10.1016/0141-0229(92)90093-4. View

14.

Chang J, El Rassi Z, Horvath C . Silica-bound polyethyleneglycol as stationary phase for separation of proteins by high-performance liquid chromatography. J Chromatogr. 1985; 319(3):396-9. DOI: 10.1016/s0021-9673(01)90578-3. View

15.

Dubois T, Jacquet A, SCHNEK A, Looze Y . The thiol proteinases from the latex of Carica papaya L. I. Fractionation, purification and preliminary characterization. Biol Chem Hoppe Seyler. 1988; 369(8):733-40. DOI: 10.1515/bchm3.1988.369.2.733. View

16.

Delgado C, Sancho P, Mendieta J, Luque J . Ligand-receptor interactions in affinity cell partitioning. Studies with transferrin covalently linked to monomethoxypoly(ethylene glycol) and rat reticulocytes. J Chromatogr. 1992; 594(1-2):97-103. DOI: 10.1016/0021-9673(92)80316-m. View

17.

Leonard M, Dellacherie E . Acylation of human hemoglobin with polyoxyethylene derivatives. Biochim Biophys Acta. 1984; 791(2):219-25. DOI: 10.1016/0167-4838(84)90012-8. View

18.

Bergstrom K, Holmberg K, Safranj A, Hoffman A, Edgell M, Kozlowski A . Reduction of fibrinogen adsorption on PEG-coated polystyrene surfaces. J Biomed Mater Res. 1992; 26(6):779-90. DOI: 10.1002/jbm.820260607. View

19.

Mole J, Horton H . Kinetics of papain-catalyzed hydrolysis of -N-benzoyl-L-arginine-p-nitroanilide. Biochemistry. 1973; 12(5):816-22. DOI: 10.1021/bi00729a005. View

20.

Rullmann J, Bellido M, Van Duijnen P . The active site of papain. All-atom study of interactions with protein matrix and solvent. J Mol Biol. 1989; 206(1):101-18. DOI: 10.1016/0022-2836(89)90527-5. View