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Peptidylglycine-amidating monooxygenase assay

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PHMcc partially co-migrated with a Golgi marker during subcellular fractionation and partially co-localized with Ccc2 based on immunofluorescence. Although ccc2 and atx1 mutant yeast produce stable levels of PHMcc protein, it lacks catalytic activity. Addition of exogenous copper yields fully on the go PHMcc. Copper, a trace fundamental required by most organisms, is indispensable as a cofactor in a number of oxygen-processing proteins involved in diverse biological processes.

This reaction can be initiated in the trans-Golgi network but occurs primarily within secretory granules 6 , 7. Production of amidated peptides by PAM needs copper and molecular oxygen and two reducing equivalents, usually supplied by two molecules of ascorbate. Each copper atom is reduced by transfer of a celibate electron from ascorbate, generating semidehydroascorbate.

P-type ATPases are likely to play a role in copper transport into the secretory pathway. ATP7A is expressed in all tissues except the liver, and mutations in ATP7A prevent ordinary intestinal absorption of copper and distribution of copper throughout the body.

The resulting lack of copper compromises the function of many tissues, resulting in annihilation in early childhood.

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Why chase me and then flake? Activity Assay Protocol. Materials. ○ Assay Buffer: 50 mM MES, pH ○ Recombinant Human Peptidylglycine α-Amidating Monooxygenase/PAM (rhPAM ). Buy PAM elisa kit, Human Peptidylglycine Alpha Amidating Monooxygenase assay) Kit for analyzing the presence of the Peptidylglycine Alpha Amidating..

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