Involved in the final step of biosynthesis of alpha-melanotropin and related biologically active peptides.the bifunctional enzyme peptidylglycine alpha-amidating monooxygenase possesses two different catalytic domains: the peptidylglycine alpha-hydroxylating monooxygenase domain and the peptidyl alpha-hydroxyglycine alpha-amidating lyase domain EC 126.96.36.199 is often called peptidylglycine alpha-amidating monooxygenase (PAM) and the alpha-amidated product is mentioned as the product of the reaction, but the alpha-amidation of glycine-extended peptides is a two-step process catalyzed by 2 enzymes: 1.Secretory vesicles were visualized by fluorescence microscope imaging in live rat atrial myocytes expressing pro ANP–enhanced green fluorescent protein (EGFP), or N-terminal–mutated fusion proteins thought to suppress the calcium-dependent aggregation of pro ANP.Results showed the following: (1) aggregates of pro ANP and coexpressed pro ANP-EGFP recruited peptidylglycine α-amidating monooxygenase (PAM)-1, an abundant atrial integral vesicle membrane protein; (2) coexpressed N-terminal–mutated (Glu23,24→Gln23,24) and N-terminal–deleted pro ANP-EGFP inhibited recruitment of PAM-1 by up to 60%; (3) 4-phenyl-3-butenoic acid (PBA) (10 μmol/L), a pharmacological inhibitor of the lumenal peptidylglycine α-hydroxylating monooxygenase domain of PAM proteins, inhibited recruitment of endogenous PAM-1 and of coexpressed pro-EGFP–PAM-1; (4) PBA had no effect on exocytosis of the potassium inward rectifier KIR2.1; (5) PBA induced a deformation of the secretory vesicles but did not inhibit docking.the system of philosophical idealism developed by George Berkeley (1685? -1753), especially his tenet that the physical world does not have an independent reality but exists as a perception of the divine mind and the flnite mind of man. The full text of this article is available online at
Peptidylglycines with a neutral amino acid residue in the penultimate position are the best substrates for the enzyme.
Accordingly, there are thousands of studies on ANP release and action, in both animal and man.
In contrast to this wealth of knowledge, the cell biology of ANP-containing secretory vesicles is far less understood.
The aim of the study was to obtain recombinant PAM and use it for insulin analogue amidation.
We stably expressed a recombinant PAM in CHO dhfr-cells in culture.