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

DOI: 10.5958/0976-0555.2015.00026.6    | Article Id: B-2528 | Page : 127-131
Citation :- Isolation and characterization of Stromelysin-3(MMP-11) like protein from a case of mammary carcinoma in a dog.Indian Journal of Animal Research.2015.(49):127-131
Sujata Sahoo*, Meena Kataria and S.K. Maiti sujatasahoo@cife.edu.in
Address : Division of Animal Biochemistry Indian Veterinary Research Institute, Izatnagar, Bareilly-243 122, India.


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.


Metastasis MMP-11 Papillary adenocarcinoma Stromelysin-3 TIMP-1.


  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.
  11. Makowski G S and Tamsby M L(1996) Calibrating gelatin zymograms with human gelatinase standards. Analytical biochem. 236:353-356
  12. 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
  13. 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
  14. 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
  15. 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
  16. Pei D, Weiss S J (1995) Furin-dependent intracellular activation of the human stromelysin-3 zymogen. Nature 375:244– 6.
  17. Rouyer N, Wolf C, Chenard M P (1994) Stromelysin-3 gene expression in human cancer: an overview. Invasion Metastasis 14:269–75
  18. 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.

Global Footprints