Structure-function studies on the biosynthesis and bioactivity of the precursor convertase PC2 and the formation of the PC2/7B2 complex.
Site directed mutagenesis of the prohormone convertase PC2 was used to define the effect of certain residues on the zymogen activation of proPC2 and on its binding to the neuroendocrine protein 7B2. These included the oxyanion hole Asp309 (D309N), the N-terminal Glu25 (E25Q and E25K) of proPC2 and the Asp519 (D519E) of the RGD motif within the P-domain of PC2. Heterologous vaccinia virus expression of the wild type and mutant PC2's in endocrine pituitary cells such as AtT20 and GH3 cells demonstrated that the most dramatic effect was observed with the D309N mutant which no longer bound pro7B2 and which exhibited a significant reduction in its capacity to produce beta-endorphin from pro-opiomelanocortin (POMC).
Benjannet
S
S
J. A. De Sève Laboratory of Molecular Neuroendocrinology, Clinical Research Institute of Montreal, Canada.
Lusson
J
J
Hamelin
J
J
Savaria
D
D
Chrétien
M
M
Seidah
N G
NG
D000818:Animals; D014408:Biomarkers, Tumor / Q000378:metabolism; D002460:Cell Line; D045683:Furin; D007163:Immunosorbent Techniques; D051379:Mice; D016297:Mutagenesis, Site-Directed; D009363:Neoplasm Proteins / Q000378:metabolism; D009419:Nerve Tissue Proteins / Q000378:metabolism*; D051859:Neuroendocrine Secretory Protein 7B2; D010907:Pituitary Hormones / Q000378:metabolism*; D011333:Pro-Opiomelanocortin / Q000378:metabolism; D045706:Proprotein Convertase 2; D043484:Proprotein Convertases; D011498:Protein Precursors / Q000378:metabolism; D012697:Serine Endopeptidases / Q000378:metabolism; D013329:Structure-Activity Relationship; D013381:Subtilisins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D014162:Transfection
D014408:Biomarkers, Tumor; D009363:Neoplasm Proteins; D009419:Nerve Tissue Proteins; D051859:Neuroendocrine Secretory Protein 7B2; D010907:Pituitary Hormones; D011498:Protein Precursors; D011333:Pro-Opiomelanocortin; C513086:Pcsk6 protein, mouse; D043484:Proprotein Convertases; D012697:Serine Endopeptidases; D013381:Subtilisins; D045683:Furin; D045706:Proprotein Convertase 2