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A biologically active angiogenesis inhibitor, human serum albumin–TIMP-2 fusion protein, secreted from Saccharomyces cerevisiae
- Source :
- Protein Expression and Purification. 53:331-338
- Publication Year :
- 2007
- Publisher :
- Elsevier BV, 2007.
-
Abstract
- Tissue inhibitor of metalloproteinase-2 (TIMP-2) is an endogenous and bi-functional inhibitor of angiogenesis. TIMP-2 is expressed in an insoluble form in Escherichia coli and secreted at a very low level from yeast. Here, we report on a high level of secretion of TIMP-2 fused with human serum albumin (HSA) from the yeast Saccharomyces cerevisiae. The secreted HSA–TIMP-2 fusion protein (87 kDa) was purified to greater than 95% homogeneity. The HSA–TIMP-2 protein inhibited approximately 81% of tube formation of human umbilical vein endothelial cells (HUVECs) when studied at a concentration of 187 μM. The systemic administration of HSA–TIMP-2 at 40 mg/kg to the C57B1/6 mouse inhibited the growth of B16BL6 tumors. Furthermore, a combination treatment of HSA–TIMP-2 with 5-fluorouracil (50 mg/kg) showed significant effects on tumor growth in this model. The high level of secretion of the biologically active angiogenesis inhibitor from S. cerevisiae should facilitate fundamental research and application studies of HSA–TIMP-2, as an attractive candidate for therapeutic agents treating angiogenesis-related diseases.
- Subjects :
- Angiogenesis
Recombinant Fusion Proteins
Genetic Vectors
Saccharomyces cerevisiae
Melanoma, Experimental
Serum albumin
Gene Expression
Angiogenesis Inhibitors
In Vitro Techniques
Matrix Metalloproteinase Inhibitors
Mice
medicine
Animals
Humans
Secretion
Cells, Cultured
Serum Albumin
Tube formation
Tissue Inhibitor of Metalloproteinase-2
biology
Endothelial Cells
biology.organism_classification
Human serum albumin
Molecular biology
Fusion protein
Angiogenesis inhibitor
Mice, Inbred C57BL
body regions
Biochemistry
embryonic structures
biology.protein
Female
Fluorouracil
Plasmids
Biotechnology
medicine.drug
Subjects
Details
- ISSN :
- 10465928
- Volume :
- 53
- Database :
- OpenAIRE
- Journal :
- Protein Expression and Purification
- Accession number :
- edsair.doi.dedup.....9cb760eca55707038b4ae43f5c867546