Banner
Current IssueEditorial BoardOnline First ArticlesArchive
Current IssueEditorial BoardOnline First ArticlesArchive

volume 49 issue 1 (february 2015) : 127-131,   Doi: 10.5958/0976-0555.2015.00026.6

Isolation and characterization of Stromelysin-3(MMP-11) like protein from a case of mammary carcinoma in a dog

S
Sujata Sahoo*
M
Meena Kataria
S
S.K. Maiti
1Division of Animal Biochemistry Indian Veterinary Research Institute, Izatnagar, Bareilly-243 122, India.
Cite article:- Sahoo* Sujata, Kataria Meena, Maiti S.K. (2025). Isolation and characterization of Stromelysin-3(MMP-11) like protein from a case of mammary carcinoma in a dog. Indian Journal of Animal Research. 49(1): 127-131. doi: 10.5958/0976-0555.2015.00026.6.

ABSTRACT

Stromelysin-3(MMP-11) is a matrix-metalloproteinase known to be over expressed in invasive tumors of human origin and suppose to have a potential role in metastasis. Dog mammary tumors are usually very invasive. Thus to characterize dog stromelysin-3, mammary tumor tissue (papillary adenocarcinoma) was used to isolate the protein by using affinity chromatography tools including Cibacron-Blue 3GA-CL Agarose, gelatin Sepharose, and  heparin Sepharose columns. Western blotting with antihuman stromelysin-3 antibody was done for identification. The result showed a heavier (120kDa) protein than the expected size of stromelysin-3. Western blot analysis of the crude sample did not show any other reactive bands. Under reducing condition, the protein size remained intact. The stromelysin-3 like protein showed mild caseinolytic activity, which increased by activators like APMA and trypsin. Whereas, its activity decreased when inhibitors like EDTA, 1, 10-Phenanthroline and TIMP-1 were used suggesting the requirement of calcium and zinc for enzyme activity just like any other metalloproteinase.

REFERENCES

  1. Andarawewa, L.K., Boulay, A, Masson, R. Mathelin, C. Stoll, I. Tomasetto, C. Chenard, C. Gintz, M. Bellocq, J. and Rio M. (2003) Dual Stromelysin- 3 Function during Natural Mouse Mammary Tumor Virus-ras Tumor Progression. Cancer Res 63: 5844-5849
  2. Basset P. Bellocq J P, Wolf C (1990) A novel matrix metalloprotease gene specifically expressed in stromal cells of breast carcinomas. Nature 348:699–704
  3. Birkadel-Hansen H, Moore W.G, Boodeen M, Windsor LJ, Birkadel-Hansen B, Decarlo A and Engler JA (1993). Matrix metalloproteinases: a review. Critial Reviews on Oral Biol. Med. 4: 197-250.
  4. Chambers A F, Matrisian L M (1997) Changing views of the role of matrix metalloproteinases in metastasis. J Natl Cancer Inst 89:1260-1270
  5. Duffy M J, McCarthy K (1998) Matrix matalloproteinases in cancer: prognostic markers and targets for therapy. Int. J. Oncol 12:1343-1348.
  6. Engel G, Heselmeyer K, Auer G, Backdahl M, Eriksson E, Linder S (1994) Correlation between stromelysin-3 mRNA level and outcome of human breast cancer. Int. J. Cancer 58: 830–835
  7. Esther E J M, Creemers J P M, Cleutjens J F M, Smits M J A P D (2001) Matrix metalloproteinase inhibition after myocardial infarction, a new approach to prevent heart failure? Circ. Res 89: 201-210
  8. Garbett E A, Reed M W R and Brown N J (1999) Proteolysis in human breast and colorectal cancer. British J. Cancer 81:287-293
  9. Huessen C and Dowdle E B (1980). Electrophoretic analysis of plasminogen activator in polyacrylamide gels containing sodium dodecylsulphate and copolymerized substrates. Analytical Biochem. 102: 196-202.
  10. Lana S E, Ogilvie G K, Hansen R A, Powers B E, Dernell, W S and Withrow S J. (2000). Identification of matrixmetalloproteinases in canine neoplastic tissue. Ameri J. Vet Res. 61:111-114.
  12. Makowski G S and Tamsby M L(1996) Calibrating gelatin zymograms with human gelatinase standards. Analytical biochem. 236:353-356
  13. Mari B P, Anderson I C, Mari S E, Ning Y, Lutz Y, Kobzik L and Shipp M A (1997) Stromelysin-3 is induced in Tumor/    Stroma co cultures and inactivated via a tumor-specific and basic fibroblast growth factordependent mechanism. J. Biol. Chem 273:618-626
  14. Murphy G, Segain J P, O’Shea M, Cockett M, Ioannou C, LefeÁ bvre O, Chambon P and Basset P (1993) The 28-kDa N-    terminal Domain of Mouse Stromelysin-3 Has the General Properties of a Weak Metalloproteinase. J. Bio. Chem, 268: 15435-15441
  15. Ogiwara K, Matsui H, Kimura A, Takahashi T (2002) Molecular cloning and partial characterization of medaka fish stromelysin-3 and its restricted expression in the oocytes of small growing follicles of the ovary. Mol Reprod Dev 61:21-31
  16. Pei D, Majmudar G, Weiss S J ( 1994) Hydrolytic inactivation of a breast carcinoma cell-derived serpin by human ST-3. J Biol Chem 269: 25849–25855
  17. Pei D, Weiss S J (1995) Furin-dependent intracellular activation of the human stromelysin-3 zymogen. Nature 375:244– 6.
  18. Rouyer N, Wolf C, Chenard M P (1994) Stromelysin-3 gene expression in human cancer: an overview. Invasion Metastasis 14:269–75
  19. Rungsipipat A, Suryasootcare B, Ousawaphlamychai L, Sailsauta A, thavawongnuwich R, Teankum K, Boomer S, Lek O, Roongroja T (2003) Neoplasm of dogs in Bangtok Thai. J. Vet Med. 33:59-66.
Published In
Indian Journal of Animal Research
Article Metrics
Metrics Icon
0
Views
0
Citations
Reviewed By
Share Article
Download Article
In this Article
    Contact us
    Follow us

    volume 49 issue 1 (february 2015) : 127-131,   Doi: 10.5958/0976-0555.2015.00026.6

    Isolation and characterization of Stromelysin-3(MMP-11) like protein from a case of mammary carcinoma in a dog

    S
    Sujata Sahoo*
    M
    Meena Kataria
    S
    S.K. Maiti
    1Division of Animal Biochemistry Indian Veterinary Research Institute, Izatnagar, Bareilly-243 122, India.
    Cite article:- Sahoo* Sujata, Kataria Meena, Maiti S.K. (2025). Isolation and characterization of Stromelysin-3(MMP-11) like protein from a case of mammary carcinoma in a dog. Indian Journal of Animal Research. 49(1): 127-131. doi: 10.5958/0976-0555.2015.00026.6.

    ABSTRACT

    Stromelysin-3(MMP-11) is a matrix-metalloproteinase known to be over expressed in invasive tumors of human origin and suppose to have a potential role in metastasis. Dog mammary tumors are usually very invasive. Thus to characterize dog stromelysin-3, mammary tumor tissue (papillary adenocarcinoma) was used to isolate the protein by using affinity chromatography tools including Cibacron-Blue 3GA-CL Agarose, gelatin Sepharose, and  heparin Sepharose columns. Western blotting with antihuman stromelysin-3 antibody was done for identification. The result showed a heavier (120kDa) protein than the expected size of stromelysin-3. Western blot analysis of the crude sample did not show any other reactive bands. Under reducing condition, the protein size remained intact. The stromelysin-3 like protein showed mild caseinolytic activity, which increased by activators like APMA and trypsin. Whereas, its activity decreased when inhibitors like EDTA, 1, 10-Phenanthroline and TIMP-1 were used suggesting the requirement of calcium and zinc for enzyme activity just like any other metalloproteinase.

    REFERENCES

    1. Andarawewa, L.K., Boulay, A, Masson, R. Mathelin, C. Stoll, I. Tomasetto, C. Chenard, C. Gintz, M. Bellocq, J. and Rio M. (2003) Dual Stromelysin- 3 Function during Natural Mouse Mammary Tumor Virus-ras Tumor Progression. Cancer Res 63: 5844-5849
    2. Basset P. Bellocq J P, Wolf C (1990) A novel matrix metalloprotease gene specifically expressed in stromal cells of breast carcinomas. Nature 348:699–704
    3. Birkadel-Hansen H, Moore W.G, Boodeen M, Windsor LJ, Birkadel-Hansen B, Decarlo A and Engler JA (1993). Matrix metalloproteinases: a review. Critial Reviews on Oral Biol. Med. 4: 197-250.
    4. Chambers A F, Matrisian L M (1997) Changing views of the role of matrix metalloproteinases in metastasis. J Natl Cancer Inst 89:1260-1270
    5. Duffy M J, McCarthy K (1998) Matrix matalloproteinases in cancer: prognostic markers and targets for therapy. Int. J. Oncol 12:1343-1348.
    6. Engel G, Heselmeyer K, Auer G, Backdahl M, Eriksson E, Linder S (1994) Correlation between stromelysin-3 mRNA level and outcome of human breast cancer. Int. J. Cancer 58: 830–835
    7. Esther E J M, Creemers J P M, Cleutjens J F M, Smits M J A P D (2001) Matrix metalloproteinase inhibition after myocardial infarction, a new approach to prevent heart failure? Circ. Res 89: 201-210
    8. Garbett E A, Reed M W R and Brown N J (1999) Proteolysis in human breast and colorectal cancer. British J. Cancer 81:287-293
    9. Huessen C and Dowdle E B (1980). Electrophoretic analysis of plasminogen activator in polyacrylamide gels containing sodium dodecylsulphate and copolymerized substrates. Analytical Biochem. 102: 196-202.
    10. Lana S E, Ogilvie G K, Hansen R A, Powers B E, Dernell, W S and Withrow S J. (2000). Identification of matrixmetalloproteinases in canine neoplastic tissue. Ameri J. Vet Res. 61:111-114.
    12. Makowski G S and Tamsby M L(1996) Calibrating gelatin zymograms with human gelatinase standards. Analytical biochem. 236:353-356
    13. Mari B P, Anderson I C, Mari S E, Ning Y, Lutz Y, Kobzik L and Shipp M A (1997) Stromelysin-3 is induced in Tumor/    Stroma co cultures and inactivated via a tumor-specific and basic fibroblast growth factordependent mechanism. J. Biol. Chem 273:618-626
    14. Murphy G, Segain J P, O’Shea M, Cockett M, Ioannou C, LefeÁ bvre O, Chambon P and Basset P (1993) The 28-kDa N-    terminal Domain of Mouse Stromelysin-3 Has the General Properties of a Weak Metalloproteinase. J. Bio. Chem, 268: 15435-15441
    15. Ogiwara K, Matsui H, Kimura A, Takahashi T (2002) Molecular cloning and partial characterization of medaka fish stromelysin-3 and its restricted expression in the oocytes of small growing follicles of the ovary. Mol Reprod Dev 61:21-31
    16. Pei D, Majmudar G, Weiss S J ( 1994) Hydrolytic inactivation of a breast carcinoma cell-derived serpin by human ST-3. J Biol Chem 269: 25849–25855
    17. Pei D, Weiss S J (1995) Furin-dependent intracellular activation of the human stromelysin-3 zymogen. Nature 375:244– 6.
    18. Rouyer N, Wolf C, Chenard M P (1994) Stromelysin-3 gene expression in human cancer: an overview. Invasion Metastasis 14:269–75
    19. Rungsipipat A, Suryasootcare B, Ousawaphlamychai L, Sailsauta A, thavawongnuwich R, Teankum K, Boomer S, Lek O, Roongroja T (2003) Neoplasm of dogs in Bangtok Thai. J. Vet Med. 33:59-66.
    In this Article
      Contact us
      Follow us
      Published In
      Indian Journal of Animal Research

      Editorial Board

      View all (0)