Expression of human prohormone convertase PC2 in a baculovirus-insect cell system.
PC2 is a member of the eukaryotic family of subtilisin-related proprotein convertases which are thought to be involved in the intracellular proteolytic processing of prohormones and proneuropeptides. The presence of only small amounts of PC2 in the secretory granules of certain mammalian neuroendocrine cell types has made the characterization and further study of this enzyme difficult. We report here the expression of proteolytically active human PC2 protein in the insect cell-baculovirus system. Human PC2 expressed in insect cells is a calcium-dependent intracellular protein active at neutral pH. In insect cells, human PC2 was found intracellularly as 75-kDa and 71-kDa proteins. Both 73-kDa and 68-kDa forms were found in the conditioned medium, but no PC2 proteolytic activity was detected. We demonstrated the presence of a soluble inhibitor in infected-cell medium which may block PC2 activity.
Fahnestock
M
M
Department of Psychiatry and Behavioural Neurosciences, McMaster University, Hamilton, Ontario, Canada.
Zhu
W
W
D000818:Animals; D016367:Baculoviridae / Q000235:genetics; D002478:Cells, Cultured; D003001:Cloning, Molecular; D005822:Genetic Vectors / Q000235:genetics; D006801:Humans; D006863:Hydrogen-Ion Concentration; D007313:Insecta / Q000166:cytology; D010447:Peptide Hydrolases / Q000096:biosynthesis / Q000235:genetics; D010455:Peptides / Q000378:metabolism; D045706:Proprotein Convertase 2; D013379:Substrate Specificity; D013381:Subtilisins / Q000037:antagonists & inhibitors / Q000096:biosynthesis / Q000235:genetics* / Q000302:isolation & purification
D010455:Peptides; D010447:Peptide Hydrolases; D013381:Subtilisins; D045706:Proprotein Convertase 2