Dissociation of collagenase-tissue inhibitor of metalloproteinases-1 (TIMP-1) complex--its application for the independent measurements of TIMP-1 and collagenase activity in crude culture media and body fluids.
Collagenase-tissue inhibitor of metalloproteinases-1 (TIMP-1) complex was prepared from activated collagenase and TIMP-1 purified from culture media of human skin fibroblasts. After having been confirmed to be a complex by zinc chelate chromatography, the complex was demonstrated to dissociate by passage through an anti-TIMP-1 monoclonal antibody-affinity column. On the basis of above evidence, a simple strategy was set up for the independent measurements of TIMP-1 concentration, and both active and total collagenase activities in crude culture media and body fluids.
Yamashita
K
K
Department of Biochemistry, School of Dentistry, Aichi-Gakuin University, Nagoya, Japan.
Zhang
J
J
Zou
L
L
Hayakawa
H
H
Noguchi
T
T
Kondo
I
I
Narita
O
O
Fujimoto
N
N
Iwata
K
K
Hayakawa
T
T
D001826:Body Fluids / Q000201:enzymology*; D002478:Cells, Cultured; D002846:Chromatography, Affinity; D017364:Collagenases / Q000032:analysis / Q000378:metabolism*; D003470:Culture Media / Q000737:chemistry*; D005260:Female; D005347:Fibroblasts / Q000201:enzymology; D005881:Gingiva / Q000201:enzymology; D006023:Glycoproteins / Q000032:analysis / Q000378:metabolism*; D006801:Humans; D008168:Lung / Q000196:embryology / Q000201:enzymology; D008297:Male; D061965:Matrix Metalloproteinase Inhibitors; D019714:Tissue Inhibitor of Metalloproteinases
D003470:Culture Media; D006023:Glycoproteins; D061965:Matrix Metalloproteinase Inhibitors; D019714:Tissue Inhibitor of Metalloproteinases; D017364:Collagenases