Release of plasminogen activator and a calcium-dependent metalloprotease from cultured sympathetic and sensory neurons.
Cultures of neurons from neonatal rat superior cervical, dorsal root, and trigeminal ganglia were grown in the absence of nonneuronal cells in serum-free defined medium. Proteins metabolically labeled with radioactive amino acids and spontaneously released into the culture medium were studied using two-dimensional gel electrophoresis and photofluorography. All three populations of neurons released 12-15 major proteins into the culture medium. Four proteins were released selectively by sympathetic neurons and two proteins were consistently released by both populations of sensory neurons but not by sympathetic neurons. Enzymatic activities are associated with at least two of the released proteins. One is a calcium-dependent metalloprotease, and the other a plasminogen activator. The calcium-dependent metalloprotease has a MW of 62 kDa, requires millimolar calcium for maximum activity, and has a restricted substrate specificity. It degraded native and denatured collagen more readily than casein, albumin, or fibronectin and denatured collagen (gelatin) was a better substrate than native collagen. The plasminogen activator released by neurons has a MW of 51 kDa and is converted to an active 32 kDa form. Its physiochemical properties are similar to urokinase and it was precipitated by a rabbit antiserum produced against human urokinase. A large fraction of both proteases was released by distal processes and/or growth cones suggesting that these proteases could be involved in growth cone functions.
Pittman
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D000818:Animals; D002118:Calcium / Q000378:metabolism*; D002154:Calpain; D003094:Collagen / Q000378:metabolism; D003546:Cysteine Endopeptidases; D010450:Endopeptidases / Q000378:metabolism*; D005353:Fibronectins / Q000378:metabolism; D005727:Ganglia, Spinal / Q000166:cytology; D005728:Ganglia, Sympathetic / Q000166:cytology; D006801:Humans; D008666:Metalloendopeptidases; D008970:Molecular Weight; D009475:Neurons, Afferent / Q000378:metabolism*; D009476:Neurons, Efferent / Q000378:metabolism*; D010960:Plasminogen Activators / Q000378:metabolism*; D011817:Rabbits; D051381:Rats; D014568:Urokinase-Type Plasminogen Activator / Q000378:metabolism
D005353:Fibronectins; D003094:Collagen; D010450:Endopeptidases; D010960:Plasminogen Activators; D014568:Urokinase-Type Plasminogen Activator; D002154:Calpain; D003546:Cysteine Endopeptidases; D008666:Metalloendopeptidases; D002118:Calcium