Skip to main content

Advertisement

SpringerLink
Log in

Variant cathepsin B activity secreted from human pancreatic cancer cell lines into protein-free chemically defined medium

  • Original Article
  • Published:
Gastroenterologia Japonica Aims and scope Submit manuscript

Summary

Cathepsin B activity was found in serum-free spent media of human pancreatic cancer cell lines, Panc-1 and MiaPaca. Cathepsin B activity was partially purified by gel filtration on TSK G3000SW, Con-A Sepharose chromatography, Phenyl-Superose column chromatography, and Mono S column chromatography. The optimal pH of cathepsin B was 7.4, and the activity was retained even at alkaline pH. Heat stability test showed tha the enzyme was heat stable; that is, 50% activity was retained after incubation at 56°C for 60 min. These results suggest that cathepsin B secreted from human pancreatic cancer cell is a variant type and may play an important role in pancreatic cancer invasion or metastasis through destruction of the surrounding extracellular matrix by its proteolytic activity.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

Abbreviations

Con A:

concanavaline A

kd:

kilodalton

Z-Phe-Phe-CHN2 :

Carbobenzoxy-L-Phenylalanyl-L-Phenylalanine CHN2

Z-Phe-Ala-CHN2 :

Carbobenzoxy-L-Phenylalanyl-L-Alanine CHN2

Z-Arg-Arg-MCA:

Carbobenzoxy-L-Arginyl-L-Arginine 4-Methyl-Coumaryl-7-Amide

Z-Phe-Arg-MCA:

Carbobenzoxy-L-Phenylalanyl-L-Arginine 4-Methyl-Coumaryl-7-Amide

Arg-MCA:

L-Arginine 4-Methyl-Coumaryl-7-Amide

Bz-Arg-MCA:

Benzoyl-L-Arginine 4-Methyl-Coumaryl-7-Amide

AMC:

7-Amino-4-Methyl-Coumarine

SDS-PAGE:

Sodium dodecyl sulfate-Polyacrylamide gel electrophoresis

References

  1. Barrett AJ, Kirschke H, Cathepsin B, Cathepsin H, and Cathepsin L. In: Methods in enzymology, Vol. 80. Academic Press, Inc. 1981;535-561

  2. Mason RW, Taylor MAJ, Etherington DJ: The purification and properties of cathepsin L from rabbit liver. Biochem J 1984;217:209–217

    PubMed  CAS  Google Scholar 

  3. Bradford M: A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principles of protein-dye binding. Anal Biochem 1976;72:248–254

    Article  PubMed  CAS  Google Scholar 

  4. Sloane BF: Lysosomal Cathepsin B: Correlation with Metastatic Potential. Science 1981;212:1151–1153

    Article  PubMed  CAS  Google Scholar 

  5. Sloane BF, Honn KV, Sadler JG, et al: Cathepsin B Activity in B 16 Melanoma Cells: A Possible Marker for Metastatic Potential. Cancer Res 1982;42:980–986

    PubMed  CAS  Google Scholar 

  6. Sloane BF, Rozhin J, Johnson K, et al: Cathepsin B: Association with plasma membrane in metastatic tumors. Proc Natl Acad Sci 1986;83:2483–2487

    Article  PubMed  CAS  Google Scholar 

  7. Mullins DE, Rohrlich ST: The role of proteinases in cellular invasiveness. Biochim Biophys Acta 1983;695:177–214

    PubMed  CAS  Google Scholar 

  8. Sloane BF, Honn KV: Cysteine proteinases and metastasis. Cancer Metastasis Reviews 1984;3:249–263

    Article  PubMed  CAS  Google Scholar 

  9. Barrett AJ: Cathepsin B and other thiol proteinases. In: Barrett AJ, ed. Proteinases in Mammalian Cells and Tissues, Elsevier/ North-Holland Biochemical Press. 1977;181-208

  10. Mort JS, Recklies AD, Poole AR: Characterization of a thiol proteinase secreted by malignant human breast tumors. Biochim Biophys Acta 1980;614:134–143

    PubMed  CAS  Google Scholar 

  11. Sloane BF, Bajkowski AS, Day NA, et al: Purification and properties of a cathepsin B-like enzyme from human tumors: comparison with cathepsin B isolated from human liver and spleen (Abstract). Proc Am Assoc Cancer Res 1984;25:4

    Google Scholar 

  12. Mason RW: Species variations amongst lysosomal cysteine proteinases. Biomed Biochim Acta 1986;45:1433–1440

    PubMed  CAS  Google Scholar 

  13. Cavanaugh PG, Sloane BF, Bajkowski A, et al: Involvement of a cathepsin B-like cysteine proteinase in platelet aggregation induced by tumor cells and their shed membrane vesicles. Clin Exp Metastasis 1983;l:297–308

    Article  Google Scholar 

  14. Recklies AD, Mort JS, Poole AR: Secretion of a thiol proteinase from mouse mammry carcinomas and its characterization. Cancer Res 1982;42:1026–1032

    PubMed  CAS  Google Scholar 

  15. Pietras RJ, Roberts JA: Cathepsin B-like enzymes. Subcellular distribution and properities in neoplastic and control cells from human ectocervix. J Biol Chem 1981;256:8536–8544

    PubMed  CAS  Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Department of Preventive Medicine, Kyoto Prefectural University of Medicine, Kawaramachi-Hirokoji, Kamigyou-ku, 602, Kyoto, Japan

    Sung min Chung & Keiichi Kawai

Authors
  1. Sung min Chung
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Keiichi Kawai
    View author publications

    You can also search for this author in PubMed Google Scholar

Rights and permissions

Reprints and permissions

About this article

Cite this article

Chung, S.m., Kawai, K. Variant cathepsin B activity secreted from human pancreatic cancer cell lines into protein-free chemically defined medium. Gastroenterol Jpn 24, 699–706 (1989). https://doi.org/10.1007/BF02774170

Download citation

  • Received:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF02774170

Key words

Search

Navigation