Purification and Characterization of Tissue-Type Plasminogen Activator in Maxillary Mucosa with Chronic Inflammation

Summary

Tissue-type plasminogen activator (TPA) was purified from maxillary mucosa with chronic inflammation and compared with urokinase. Purification procedure consisted of the extraction from delipidated mucosa with 0.3M potassium acetate buffer (pH 4.2), 66% saturation of ammonium sulfate, zinc chelate-Sepharose, concanavalin A-Sepharose and Sephadex G-100 gel filtration chromatographies.

The molecular weight of the TPA was approximately 58,000 ± 3,000. Its activity was enhanced in the presence of fibrin and was quenched by placental urokinase inhibitor, but not quenched by anti-urokinase antibody. The TPA made no precipitin line against anti-urokinase antibody, while urokinase did.

All these findings indicate that the TPA in maxillary mucosa with chronic inflammation is immunologically dissimilar to urokinase and in its affinity for fibrin.

Keywords

Chronic sinusitis - Maxillary mucosa - Plasminogen activator - Urokinase

• References

• 1 Astrup T. Fibrinolysis in the organism. Br J Hematol 1956; 9: 781-806
• 2 Bachmann F, Fletcher AP, Alkjaersig N, Sherry S. Partial purification and properties of the plasminogen activator from pig heart. Biochemistry 1964; 3: 1578-1585
• 3 Rijken DC, Wijngaards G, Zaal-De Jong M, Welbergen J. Purification and partial characterization of plasminogen activator from human uterine tissues. Biochim Biophys Acta 1979; 580: 140-153
• 4 Kok P, Astrup T. Isolation and purification of a tissue plasminogen activator and its comparison with urokinase. Biochemistry 1969; 8: 79-85
• 5 Rijken DC, Wijngaards G, Welbergen J. Immunological characterization of plasminogen activator activities in human tissues and body fluids. J Lab Clin Med 1981; 97: 477-486
• 6 Hamaguchi Y. Partial purification and properties of tissue-type plasminogen activator from nasal and maxillary mucosae. Mie Med J 1983; 33: 57-67
• 7 Hamaguchi Y, Sakakura Y, Majima Y, Ohi M, Mitsui H, Harada T. Kinetics of immunoglobulin in nasal secretions of chronic sinusitis and nasal allergy. Practica Otologica 1981; 74: 1170-1181
• 8 Hjelm H, Hjelm K, Sjoquist J. Protein A from staphylococcus aureus. Its isolation by affinity chromatography and its use as an immunosorbent for isolation of immunoglobulins. FEBS Letts 1972; 28: 73-76
• 9 Porath J, Carlsson J, Olsson I. Metal chelate affinity chromatography, a new approach to protein fractionation. Nature 1975; 258: 598-599
• 10 Morita T, Kato S, Iwanaga S, Takada K, Kimura T, Sakakibara S. New fluorogenic substrates for α-thrombin, factor X_a, kallikrein and urokinase. J Biochem 1977; 82: 1495-1499
• 11 Kato H, Adachi N, Ohno Y, Iwanaga S, Takada K, Sakakibara S. New fluorogenic peptide substrate for plasmin. J Biochem 1980; 88: 183-190
• 12 Kanaoka Y, Takahashi T, Nakayama H, Takada K, Sakakibara S. Synthesis of a key fluorogenic amide. Fluorescence assays for trypsin and papain. Chem Pharm Bull 1977; 25: 3126-3128
• 13 Kato H, Adachi N, Iwanaga S, Abe K, Takada K, Kimura T, Sakakibara S. A new fluorogenic substrate method for estimation of kallikrein in urine. J Biochem 1980; 87: 1127-1132
• 14 Astrup T, Müllertz S. The fibrin plate method for estimating fibrinolytic activity. Arch Biochem 1952; 40: 346-351
• 15 Deutsch D, Mertz E. Plasminogen purification from human plasma by affinity chromatography. Science 1970; 170: 1095-1096
• 16 Cole ER, Bachmann F. Purification and properties of a plasminogen activator from pig heart. J Biol Chem 1977; 252: 3729-3737
• 17 Lijnen HR, Collen D. Interaction of plasminogen activators and inhibitors with plasminogen and fibrin. Semin Thromb 1982; 8: 2-10