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.
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.
Institut für Biochemie der RWTH Aachen, Germany.
P
P
Kurschat
L
L
Graeve
A
A
Erren
P
P
Gatsios
S
S
Rose-John
E
E
Roeb
H
H
Tschesche
A
A
Koj
P C
PC
Heinrich
D000595:Amino Acid Sequence; D000818:Animals; D016367:Baculoviridae / Q000235:genetics; D002478:Cells, Cultured; D006023:Glycoproteins / Q000096:biosynthesis / Q000302:isolation & purification*; D006031:Glycosylation; D008297:Male; D061965:Matrix Metalloproteinase Inhibitors; D008657:Metabolic Clearance Rate; D051379:Mice; D008969:Molecular Sequence Data; D011480:Protease Inhibitors / Q000302:isolation & purification*; D051381:Rats; D017207:Rats, Sprague-Dawley; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000302:isolation & purification; D018411:Spodoptera; D014018:Tissue Distribution; D019714:Tissue Inhibitor of Metalloproteinases
D006023:Glycoproteins; D061965:Matrix Metalloproteinase Inhibitors; D011480:Protease Inhibitors; D011994:Recombinant Proteins; D019714:Tissue Inhibitor of Metalloproteinases