TY - JOUR
T1 - Localization of protease nexin‐1 on the fibroblast extracellular matrix
AU - Farrell, David H.
AU - Wagner, Steven L.
AU - Yuan, Robert H.
AU - Cunningham, Dennis D.
PY - 1988/2
Y1 - 1988/2
N2 - Protease nexin‐1 (PN‐1) is a protease inhibitor that is secreted by fibroblasts and several other cultured cells. PN‐1 forms complexes with certain serine proteases in the extracellular environment including thrombin, urokinase, and plasmin. The complexes then bind to the cells and are rapidly internalized and degraded. This report demonstrates that PN‐1 is present on the surface of fibroblasts, bound to the extracellular matrix. Immunofluorescent studies showed that PN‐1 colocalized with fibronectin on both intact cells and in preparations of extracellular matrix made from these cells. In contrast, PN‐1 did not colocalize with the epidermal growth factor receptor, a plasma membrane marker. An enzyme‐lined immunosorbent assay was developed which showed that the extracellular matrix contained at least 60–80% of the cellular immunoreactive PN‐1. Extraction of the matrix with 2 M NaCl removed PN‐1 in a form which reacted with 125l‐thrombin to form complexes which were immunoprecipitated by anti‐PN‐1 lgG and were of identical size as complexes made from soluble PN‐1 and 125l‐thrombin. These data indicate that in addition to its role as a soluble protease inhibitor, PN‐1 is also a component of the extracellular matrix and might control its proteolysis.
AB - Protease nexin‐1 (PN‐1) is a protease inhibitor that is secreted by fibroblasts and several other cultured cells. PN‐1 forms complexes with certain serine proteases in the extracellular environment including thrombin, urokinase, and plasmin. The complexes then bind to the cells and are rapidly internalized and degraded. This report demonstrates that PN‐1 is present on the surface of fibroblasts, bound to the extracellular matrix. Immunofluorescent studies showed that PN‐1 colocalized with fibronectin on both intact cells and in preparations of extracellular matrix made from these cells. In contrast, PN‐1 did not colocalize with the epidermal growth factor receptor, a plasma membrane marker. An enzyme‐lined immunosorbent assay was developed which showed that the extracellular matrix contained at least 60–80% of the cellular immunoreactive PN‐1. Extraction of the matrix with 2 M NaCl removed PN‐1 in a form which reacted with 125l‐thrombin to form complexes which were immunoprecipitated by anti‐PN‐1 lgG and were of identical size as complexes made from soluble PN‐1 and 125l‐thrombin. These data indicate that in addition to its role as a soluble protease inhibitor, PN‐1 is also a component of the extracellular matrix and might control its proteolysis.
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U2 - 10.1002/jcp.1041340203
DO - 10.1002/jcp.1041340203
M3 - Article
C2 - 3279057
AN - SCOPUS:0023838288
SN - 0021-9541
VL - 134
SP - 179
EP - 188
JO - Journal of Cellular Physiology
JF - Journal of Cellular Physiology
IS - 2
ER -