Antiproteolytic activity of goose pancreas: Purification, inhibitory properties and amino-acid sequence of a Kazal type trypsin inhibitor

• Autorzy: Wilimowska-Pelc A. , Stachowiak D. , Gladysz M. , Olichwier Z. , Polanowski A.
• Języki publikacji: EN

Abstrakty

EN

A trypsin inhibitor of Kazal type has been isolated from goose pancreas by affinity chromatography on immobilized anhydrotrypsin, anion exchange and reverse phase HPLC. It inhibits bovine β-trypsin with the association constant (Ka) of 5.99 x 10 M-1. The complete amino-acid sequence was determined following CNBr treatment. The protein comprised a total of 69 amino-acid residues, corresponding to a molecular mass of 7.7 kDa. The P1-P'1 reactive site bond of the inhibitor was localized at position Lys25-Met26. The amino-acid sequence of GPTI shows extremely high homology to that of other inhibitors isolated from pancreas of birds.

Słowa kluczowe

EN

anhydrotrypsin; affinity chromatography; bovine beta-trypsin; trypsin inhibitor; goose pancreas; purification; amino acid sequence; antiproteolytic activity

• Wydawca: -
• Czasopismo: Acta Biochimica Polonica
• Rocznik: 1996
• Tom: 43
• Numer: 3
• Opis fizyczny: p.489-496,fig.

Twórcy

autor

Wilimowska-Pelc A.

• University of Wroclaw, Tamka 2, 50-137 Wroclaw, Poland

autor

Stachowiak D.

autor

Gladysz M.

autor

Olichwier Z.

autor

Polanowski A.

Bibliografia

• 1. Kazal, L.A., Spicer, D.S. & Brahinsky, R.A. (1948) Isolation of a crystalline trypsin inhi­bitor-anticoagulant protein from pancreas. ). Am. Chem. Soc. 70, 3034-3040.
• 2. Bartelt, D.C., Shapanaka, R. & Green, L.J. (1977) The primary structure of the human pancreatic secretory trypsin inhibitor. Arch. Biochem. Bio- phys.179,189-199.
• 3. Green, L.J. & Bartelt, D.C. (1969) The structure of the bovine pancreatic secretory trypsin inhibitor — Kazal's inhibitor. II. The order of the tryptic peptides. J. Biol. Chem.244, 2646-2657.
• 4. Bartelt, D.C. & Green, L.J. (1971) The primary structure of the porcine pancreatic secretory trypsin inhibitor I. J. Biol. Chem.246, 2218-2229.
• 5. Uda, K., Ogawa, M., Shibata, T., Murata, A., Mori, T., Kikuchi, N., Yoshida, N., Tsunasawa, S. & Saiyama, F. (1988) Purification, characte­rization and amino-acid sequencing of two pancreatic secretory trypsin inhibitors in rat pancreatic juice. Biol. Chem. Hoppe-Seyler369 (Suppl.) 55-61.
• 6. Conlon, M.J., Kim, Ch.B. & Magee, F.D. (1990) Isolation and structural characterization of a molecular variant of dog pancreatic secretory trypsin inhibitor. Int. J. Pancreatol.8,59-64.
• 7. Laskowski, M. & Wu, F.C. (1953) Temporary inhibition of trypsin. /. Bid. Chem.204, 797-805.
• 8. Ogawa, M. (1988) Pancreatic secretory trypsin inhibitor as an acute phase reactant. Clin. Biochem.21,19-25.
• 9. Pubols, M.H. (1990) Isolation, purification and amino acid sequence of a secretory trypsin inhibitor from the chicken pancreas. Poultry Sci. , 640-646.
• 10. Laskowski, M., Jr., Kato, I., Kohr, W.J., March, C.J. & Bogard, W.C. (1980) Evolution of the family of serine proteinase inhibitors homo­logous to pancreatic secretory trypsin inhibitor (Kazal). Protides Biol. Fluids Proc. Colloq.28, 123-128.
• 11. Wilimowska-Pelc, A. & Mejbaum-Katzenellen- bogen, W. (1978) A simple method for isolating trypsin from trichloroacetic acid extracts of bovine pancreas. Anal. Biochem.90,816-820.
• 12. Liepniecks, J.J. & light, A. (1974) Preparation of 3-trypsin by affinity chromatography of entero- kinase activated bovine trypsinogen. Anal. Biochem.60, 395-404.
• 13. Ako, H., Foster, R.J. & Ryan, C.A. (1972) The preparation of anhydro-trypsin and its reacti­vity with naturally occurring proteinase inhi­bitors. Biochem. Biophys. Res. Commun.47, 1402-1407.
• 14. March, S.C., Parikh, I. & Cutrecasas, P. (1974) A simplified method for cyanogen bromide acti­vation of agarose for affinity chromatography. Anal. Biochem.60,149-152.
• 15. Wctforek, W., Polanowski, A. & Wilusz, T. (1996) The use of sequential affinity chromatography for separation of human neutrophile elastase, cathepsin G and azurocidin. Acta Biochim. Polon. , 503-506.
• 16. Goa, J. (1953) A microbiuret method for protein determination. Determination of total protein in cerebrospinal fluid. Scand. J. Clin. Lab. Invest. 5, 218-222.
• 17. Chase, T. & Shaw, E. (1970) Titration of trypsin, plasmin and thrombin with p-nitrophenyl p-gu- anidinobenzoate HCl. Methods Enzymol.19, 20-27.
• 18. Erlanger, B.F., Kokowsky, N. & Cohen, W. (1961) The preparation and properties of two newchromogenie of trypsin. Arch. Diochem. Biophys.95,271-278.
• 19. Nakajima, K., Powers, J.C., Asche, B. & ­merman M. (1979) the extended sub­strate binding site of cathepsin G and human leucocyte elastase. /. Biol. Chem.254,4027-4032.
• 20. Empie, M.W. & Laskowski, M., Jr. (1982) and kinetics of single residue replacements in avian ovomucoid third do­mains: Effect in inhibitor interaction with serine proteinases. Biochemistry21,2274-2284.
• 21. Otlewski, J., Zbyryt, ., Krokoszyńska, . & , T. (1990) of serine proteinases by squash inhibitors. Biol. Chem. Hoppe-Seyler 371, 589-594.
• 22. Laemmli, U. (1970) of structural pro­teins during assembly of the head of bacteriophage T4. Nature (London)227, 680-685.
• 23. Davis, B.J. (1964) electrophoresis II. Method and application to human serum proteins. Ann. N.Y. Acad. Sci. U.S.A.121, 404-427.
• 24. Polanowski, A., Wilusz, T., Blum, MS., Esco- ubas, P., Schmidt, J.O. & , J. (1992) proteinase inhibitor profiles in the hemolymph of a wide range of insect species. Comp. Biochem. Physiol.102 , 757-760.
• 25. Frankel-Conrat, H. (1957) for inves­tigating the essential groups for enzyme activity. Methods Enzymol.4,247-269.
• 26. Cohen, S.A. & , D.P. (1993) of a fluorescent derivatizing reagent, 6-amino- quinolyl-N-hydroxysuccinimidyl carbamate and its application for the analysis of hydro- lysate amino acids viahigh-performance liquid chromatography. Anal. Biochem.211,279-287.