Endothelial and serum factors which include apolipoprotein A1 tether elastin to smooth muscle cells inducing serine elastase activity via tyrosine kinase-mediated transcription and translation.
We previously reported that serine elastase activity is induced in cultured porcine pulmonary artery (PA) smooth muscle cells (SMC) following serum stimulation by a mechanism involving adhesion of elastin to an elastin binding protein and tyrosine kinase activity. The present study demonstrates that a PA endothelial cell factor also promotes a fourfold increase in elastin adhesion to PA SMC and a twofold increase in serine elastase activity. The mechanism involves tethering of the factor to SMC, since [3H]-elastin pre-incubated with serum or endothelial cell (EC)-conditioned medium or SMC pre-treated with serum accelerates binding of elastin and tyrosine-kinase related elastase activity. The serum factor appears to interact with integrins as elastase induction is partially inhibited by RGD peptides. The elastase-inducing properties of serum could not, however, be attributed to several RGD-containing proteins. While a 120 kD fibronectin fragment partially reproduced the effect, it was not found in the serum fraction containing elastase-inducing activity. Instead, a 27 kD serum protein was enriched by elastin affinity chromatography, identified as apolipoprotein (Apo) A1 by microsequence analysis, and found to have about 50% of the elastase-inducing activity of serum. Elastase induction is inhibited by actinomycin and cycloheximide, suggesting a requirement for mRNA transcription and protein synthesis. Our results suggest a novel cell-extracellular matrix interaction whereby a soluble factor, in this case a lipoprotein, binds and tethers a matrix component to the cell surface and induces tyrosine kinase-dependent transcription of mRNA culminating in substrate proteolysis.
Thompson
K
K
Division of Cardiovascular Research, Hospital for Sick Children, Toronto, Ontario, Canada.
Kobayashi
J
J
Childs
T
T
Wigle
D
D
Rabinovitch
M
M
D000818:Animals; D016632:Apolipoprotein A-I / Q000378:metabolism*; D001665:Binding Sites; D001798:Blood Proteins / Q000378:metabolism; D002478:Cells, Cultured; D004549:Elastin / Q000378:metabolism*; D004730:Endothelium, Vascular / Q000378:metabolism; D004789:Enzyme Activation; D009131:Muscle, Smooth, Vascular / Q000378:metabolism*; D010196:Pancreatic Elastase / Q000378:metabolism*; D011485:Protein Binding; D015398:Signal Transduction; D013552:Swine
D016632:Apolipoprotein A-I; D001798:Blood Proteins; D004549:Elastin; D010196:Pancreatic Elastase