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Expression of a biologically active murine tissue inhibitor of metalloproteinases-1 (TIMP-1) in baculovirus-infected insect cells. Purification and tissue distribution in the rat.

作者信息

Kurschat P, Graeve L, Erren A, Gatsios P, Rose-John S, Roeb E, Tschesche H, Koj A, Heinrich P C

机构信息

Institut für Biochemie der RWTH Aachen, Germany.

出版信息

Eur J Biochem. 1995 Dec 1;234(2):485-91. doi: 10.1111/j.1432-1033.1995.485_b.x.

Abstract

Murine tissue inhibitor of metalloproteinases-1 (mTIMP-1) was expressed in baculovirus-infected insect cells (Sf9). The protein secreted into the culture medium was purified to homogeneity by means of heparin-Sepharose CL-6B and FPLC. The purified protein showed metalloproteinase-inhibitory activity in two independent assays: reverse zymography and inhibition of collagenase activity. Digestion of the recombinant TIMP-1 with peptide-N-glycanaseF revealed that both N-glycosylation sites are used. 125I-mTIMP-1 intraveneously injected into a male Sprague Dawley rat disappeared within 2 min from the circulation. 5 min after injection more than 50% of the 125I-mTIMP-1 were found in the liver and 20% in the kidneys. At later times, trichloroacetic-acid-soluble material accumulated in the intestinal tract.

摘要

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