A biologically active angiogenesis inhibitor, human serum albumin-TIMP-2 fusion protein, secreted from Saccharomyces cerevisiae.
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 (87kDa) 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 187microM. The systemic administration of HSA-TIMP-2 at 40mg/kg to the C57B1/6 mouse inhibited the growth of B16BL6 tumors. Furthermore, a combination treatment of HSA-TIMP-2 with 5-fluorouracil (50mg/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.
Kang
Woo Kyu
WK
School of Bioscience and Biotechnology, Chungnam National University, Daejeon 305-764, Republic of Korea.
Park
Eun-Kyu
EK
Lee
Hee Suk
HS
Park
Byung-Young
BY
Chang
Jae-Young
JY
Kim
Min-Young
MY
Kang
Hyun Ah
HA
Kim
Jeong-Yoon
JY
D020533:Angiogenesis Inhibitors / Q000096:biosynthesis* / Q000235:genetics* / Q000302:isolation & purification / Q000494:pharmacology; D000818:Animals; D002478:Cells, Cultured; D042783:Endothelial Cells / Q000187:drug effects; D005260:Female; D005472:Fluorouracil / Q000494:pharmacology; D015870:Gene Expression; D005822:Genetic Vectors; D006801:Humans; D066298:In Vitro Techniques; D061965:Matrix Metalloproteinase Inhibitors; D008546:Melanoma, Experimental / Q000188:drug therapy / Q000473:pathology; D051379:Mice; D008810:Mice, Inbred C57BL; D010957:Plasmids / Q000235:genetics; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis / Q000235:genetics / Q000302:isolation & purification / Q000494:pharmacology; D012441:Saccharomyces cerevisiae / Q000235:genetics*; D012709:Serum Albumin / Q000096:biosynthesis* / Q000235:genetics* / Q000302:isolation & purification / Q000494:pharmacology; D019716:Tissue Inhibitor of Metalloproteinase-2 / Q000096:biosynthesis* / Q000235:genetics* / Q000302:isolation & purification / Q000494:pharmacology
D020533:Angiogenesis Inhibitors; D061965:Matrix Metalloproteinase Inhibitors; D011993:Recombinant Fusion Proteins; D012709:Serum Albumin; D019716:Tissue Inhibitor of Metalloproteinase-2; D005472:Fluorouracil