Molecular Cloning and Synthesis of Biologically Active Human Tissue Inhibitor of Metalloproteinases in Yeast

Tissue inhibitor of metalloproteinases (TIMP) is a widely distributed glycoprotein that stochiometrically inactivates metalloproteinases involved in connective tissue catabolism. Here we report the cDNA cloning of TIMP from human fibroblastic MRC5 cells using a single 42–base oligonucleotide probe. Expression in S. cerevisiae of complete TIMP cDNA yielded insoluble protein aggregates. Biologically active TIMP was reconstituted from the yeast product by a denaturation/renaturation procedure.