Identification and localisation of a synaptosomal membrane prolyl endopeptidase from bovine brain.
Prolyl endopeptidase, which has long been recognised for its importance in the degradation of several neuropeptides such as thyroliberin, luteinising hormone releasing hormone, angiotensin, substance P and neurotensin, has been widely characterised as a cytosolic enzyme. However, in this paper, we report the presence of a prolyl endopeptidase activity in the particulate fractions of bovine brain, which is distinct from that in the cytoplasm. This previously uncharacterised activity was found to reside in the synaptosomal membranes, a location which is highly significant for the inactivation of neuropeptides in brain. Following vigorous salt washing and osmotic shock, the prolyl endopeptidase activity was released from the membranes by treatment with the detergent Triton X-100, and was partially purified by gel filtration on a Sephacryl S-200HR column. This prolyl endopeptidase activity was shown to have a molecular mass (87 kDa) higher than the cytosolic prolyl endopeptidase but, from initial investigation, appears to demonstrate a similarly broad substrate specificity towards proline-containing neuropeptides. The partially purified enzyme was inhibited by certain thiol-protease inhibitors and was also found to be sensitive to the metal chelator 1,10-phenanthroline.
School of Biological Sciences, Dublin City University, Ireland.
R M
RM
O'Leary
B
B
O'Connor
D000818:Animals; D001921:Brain / Q000201:enzymology*; D002417:Cattle; D002850:Chromatography, Gel; D002851:Chromatography, High Pressure Liquid; D007202:Indicators and Reagents; D007425:Intracellular Membranes / Q000201:enzymology; D000085822:Prolyl Oligopeptidases; D012492:Salts; D012697:Serine Endopeptidases / Q000302:isolation & purification / Q000378:metabolism*; D015842:Serine Proteinase Inhibitors / Q000494:pharmacology; D013347:Subcellular Fractions / Q000201:enzymology; D013379:Substrate Specificity; D013574:Synaptosomes / Q000201:enzymology*; D013973:Thyrotropin-Releasing Hormone / Q000378:metabolism
D007202:Indicators and Reagents; D012492:Salts; D015842:Serine Proteinase Inhibitors; D013973:Thyrotropin-Releasing Hormone; D012697:Serine Endopeptidases; D000085822:Prolyl Oligopeptidases