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Literature summary for 1.14.17.3 extracted from

  • Chauhan, S.; Kline, C.D.; Mayfield, M.; Blackburn, N.J.
    Binding of copper and silver to single-site variants of peptidylglycine monooxygenase reveals the structure and chemistry of the individual metal centers (2014), Biochemistry, 53, 1069-1080 .
    View publication on PubMedView publication on EuropePMC
Search for more literature for 1.14.17.3

Protein Variants

Protein Variants Comment Organism
H107A site-directed mutagenesis, altered reaction with CO compared to wild-type Rattus norvegicus
H107A/H108A site-directed mutagenesis, removal of two of three histidines prevents metal binding at the H-center, the double His mutant H107H108A binds copper only in the M-site and therefore contains about 1 equivalent copper per protein. The PHM variant, when metalated, binds copper and silver at only a single center Rattus norvegicus
H108A site-directed mutagenesis, altered reaction with CO compared to wild-type Rattus norvegicus
H242A site-directed mutagenesis, the CuM site mutant which has an empty M site in the reduced state, does not react with CO in the presence or absence of peptide substrate. The PHM variant, when metalated, binds copper and silver at only a single center. The H242A variant is an M-site deletion mutant that removes one of the two histidines necessary for tight binding of copper to the M-site Rattus norvegicus
M109I site-directed mutagenesis, altered reaction with CO compared to wild-type Rattus norvegicus

Metals/Ions

Metals/Ions Comment Organism Structure
Cu2+ a pair of uncoupled copper centers separated by 11 A termed CuH and CuM, an unusual Cu(I)S-Met interaction at the oxygen binding M-site, and the postulated Cu(II)-superoxo intermediate, structure and chemistry of the individual metal centers. The copper centers are involved in the catalytic mechanism, the M-center binds two histidines and a methionine at all pHs, while the H-center is two-coordinate at neutral pH but coordinated another methionine S ligand at low pH. Analysis of CO complex formations of wild-type and mutant enzymes at pH 3.5-7.5, detailed overview Rattus norvegicus
additional information Ag(I) is iso-electronic with Cu(I) and has been shown to be handled by similar cellular transport processes. Although complexes of Ag in the Ag(III) state exists, the instability of the Ag(II) state renders the metal unsuitable for a functional substitute for copper in enzymes such as PHM. When enzyme mutants H107A/H108A and M109I (a wild-type analogue with both copper sites intact) are incubated with excess AgNO3, each variant binds a single Ag(I) ion, from which it is inferred that Ag(I) binds selectively at the M-center with little or no affinity for the H-center Rattus norvegicus

Organism

Organism UniProt Comment Textmining
Rattus norvegicus P14925
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-

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
diiodotyrosylglycine + ascorbate + O2
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 Rattus norvegicus diiodotyrosyl(2-hydroxyglycine) + dehydroascorbate + H2O
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 ?

Synonyms

Synonyms Comment Organism
PHM
-
 Rattus norvegicus

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
37
-
 assay at Rattus norvegicus

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
5.5
-
 assay at Rattus norvegicus

Cofactor

Cofactor Comment Organism Structure
ascorbate
-
 Rattus norvegicus

General Information

General Information Comment Organism
additional information structure of the PHM enzyme active site taken from PDB ID 1OPM structure with bound substrate diiodotyrosylglycine: the H-site coordinates to the Ndelta atom of H107, H108, and Nepsilon of H172, and the oxygen binding (catalytic) M-site coordinates to Nepsilon of H242 and H244 and the thiother S of M314. The side chain of the conserved E313 forms an H-bond to the main chain amide nitrogen of H244 Rattus norvegicus