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Literature summary for 1.14.17.3 extracted from

  • Zielinski, M.; Wojtowicz-Krawiec, A.; Mikiewicz, D.; Kesik-Brodacka, M.; Cecuda-Adamczewska, V.; Marciniak-Rusek, A.; Sokolowska, I.; Lukasiewicz, N.; Gurba, L.; Odrowaz-Sypniewski, M.; Baran, P.; Plucienniczak, G.; Plucienniczak, A.; Borowicz, P.; Szewczyk, B.
    Expression of recombinant human bifunctional peptidylglycine alpha-amidating monooxygenase in CHO cells and its use for insulin analogue modification (2016), Protein Expr. Purif., 119, 102-109 .
    View publication on PubMed
Search for more literature for 1.14.17.3

Application

Application Comment Organism
synthesis usage of the recombinant PAM for insulin analogue amidation producing insulin glargine amide, an insulin derivative that shows a time/effect profile which is distinctly more flat and thus more advantageous than insulin glargine itself. The enzyme is used to modify glycine-extended A22(G)-B31(K)-B32(R) human insulin analogue (GKR). Hypoglycemic activity of amidated and non-amidated insulin is compared Homo sapiens

Cloned(Commentary)

Cloned (Comment) Organism
gene PAM, stable recombinant expression of PAM in CHO dhfr cells, construction of pCG/dhfr plasmid with added PAMLID gene incorporating the altered leader sequence Homo sapiens

Metals/Ions

Metals/Ions Comment Organism Structure
Cu2+ dependent on Homo sapiens

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
insulin + ascorbate + O2 Homo sapiens
-
 ? + dehydroascorbate + H2O
-
 ?

Organism

Organism UniProt Comment Textmining
Homo sapiens P19021
-
-

Purification (Commentary)

Purification (Comment) Organism
recombinant PAM from CHO dhfr cells by ammonium sulfate fractionation and anion exchange chromatography Homo sapiens

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
insulin + ascorbate + O2
-
 Homo sapiens ? + dehydroascorbate + H2O
-
 ?
insulin glargine + ascorbate + O2
-
 Homo sapiens ? + dehydroascorbate + H2O
-
 ?

Synonyms

Synonyms Comment Organism
PAM
-
 Homo sapiens
peptidylglycine alpha-amidating monooxygenase
-
 Homo sapiens

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
37
-
 assay at Homo sapiens

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
6
-
 assay at Homo sapiens

Cofactor

Cofactor Comment Organism Structure
ascorbate
-
 Homo sapiens

General Information

General Information Comment Organism
physiological function PAM is the only known enzyme responsible for the bioconversion of glycine C-terminal prohormones into des-glycine alpha-amidated products and glyoxylate. PAM is a bifunctional, type-II copper monooxygenase that consists of two independent catalytic domains, PHM and PAL. The PAM reaction is a two-step process. Initially, PHM removes the pro-S hydrogen to allow hydroxylation of the alpha-glycyl carbon, resulting in an alpha-hydroxylated intermediate. PHM is a molecular oxygen, Cu(II) and ascorbate (reductant) dependent enzyme. The second catalytic domain, PAL, dealkylates the alpha-hydroxylated intermediate, yielding the alpha-amidated product and glyoxylate. PAM is responsible for the posttranslational modification of many important neuropeptides, including oxytocin, vasopressin, ACTH, alphaMSH, VIP, substance P, neuropeptide Y, cholecystokinin, gastrin, and a large number of other molecules. Hypoglycemic effect of the alpha-amidated analogue of recombinant human insulin Homo sapiens