Coordination of peroxide to the CuM center of peptidylglycine α-hydroxylating monooxygenase (PHM): Structural and computational study
Many bioactive peptides, such as hormones and neuropeptides, require amidation at the C terminus for their full biological activity. Peptidylglycine α-hydroxylating monooxygenase (PHM) performs the first step of the amidation reaction - the hydroxylation of peptidylglycine substrates at the Cα posit...
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2013
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| Acceso en línea: | https://bibliotecadigital.exactas.uba.ar/collection/paper/document/paper_09498257_v18_n2_p223_Rudzka http://hdl.handle.net/20.500.12110/paper_09498257_v18_n2_p223_Rudzka |
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paper:paper_09498257_v18_n2_p223_Rudzka2023-06-08T15:54:05Z Coordination of peroxide to the CuM center of peptidylglycine α-hydroxylating monooxygenase (PHM): Structural and computational study Amidation of peptides Copper-containing proteins Peptidylglycine α-hydroxylating monooxygenase Peroxide copper H copper ion copper M cupric ion cuprous ion glycine hydrogen peroxide oxygen derivative oxygenase peptidylglycine alpha hydroxylating monooxygenase peroxide unclassified drug amidation animal cell article carboxy terminal sequence catalysis complex formation controlled study crystal structure energy enzyme active site enzyme binding enzyme metabolism female hamster hydroxylation isomer ligand binding molecular interaction molecular mechanics molecular model nonhuman priority journal quantum mechanics structure activity relation structure analysis X ray diffraction Animals Catalytic Domain CHO Cells Computer Simulation Coordination Complexes Copper Cricetinae Crystallography, X-Ray Hydrogen Bonding Hydrogen Peroxide Mixed Function Oxygenases Models, Molecular Multienzyme Complexes Quantum Theory Rats Many bioactive peptides, such as hormones and neuropeptides, require amidation at the C terminus for their full biological activity. Peptidylglycine α-hydroxylating monooxygenase (PHM) performs the first step of the amidation reaction - the hydroxylation of peptidylglycine substrates at the Cα position of the terminal glycine. The hydroxylation reaction is copper- and O2-dependent and requires 2 equiv of exogenous reductant. The proposed mechanism suggests that O2 is reduced by two electrons, each provided by one of two nonequivalent copper sites in PHM (CuH and CuM). The characteristics of the reduced oxygen species in the PHM reaction and the identity of the reactive intermediate remain uncertain. To further investigate the nature of the key intermediates in the PHM cycle, we determined the structure of the oxidized form of PHM complexed with hydrogen peroxide. In this 1.98-Å-resolution structure (hydro)peroxide binds solely to CuM in a slightly asymmetric side-on mode. The O-O interatomic distance of the copper-bound ligand is 1.5 Å, characteristic of peroxide/hydroperoxide species, and the Cu-O distances are 2.0 and 2.1 Å. Density functional theory calculations using the first coordination sphere of the CuM active site as a model system show that the computed energies of the side-on L3CuM(II)-O2 2- species and its isomeric, end-on structure L3CuM(I)-O 2 ·- are similar, suggesting that both these intermediates are significantly populated within the protein environment. This observation has important mechanistic implications. The geometry of the observed side-on coordinated peroxide ligand in L3CuM(II)O 2 2- is in good agreement with the results of a hybrid quantum mechanical-molecular mechanical optimization of this species. © 2012 SBIC. 2013 https://bibliotecadigital.exactas.uba.ar/collection/paper/document/paper_09498257_v18_n2_p223_Rudzka http://hdl.handle.net/20.500.12110/paper_09498257_v18_n2_p223_Rudzka |
| institution |
Universidad de Buenos Aires |
| institution_str |
I-28 |
| repository_str |
R-134 |
| collection |
Biblioteca Digital - Facultad de Ciencias Exactas y Naturales (UBA) |
| topic |
Amidation of peptides Copper-containing proteins Peptidylglycine α-hydroxylating monooxygenase Peroxide copper H copper ion copper M cupric ion cuprous ion glycine hydrogen peroxide oxygen derivative oxygenase peptidylglycine alpha hydroxylating monooxygenase peroxide unclassified drug amidation animal cell article carboxy terminal sequence catalysis complex formation controlled study crystal structure energy enzyme active site enzyme binding enzyme metabolism female hamster hydroxylation isomer ligand binding molecular interaction molecular mechanics molecular model nonhuman priority journal quantum mechanics structure activity relation structure analysis X ray diffraction Animals Catalytic Domain CHO Cells Computer Simulation Coordination Complexes Copper Cricetinae Crystallography, X-Ray Hydrogen Bonding Hydrogen Peroxide Mixed Function Oxygenases Models, Molecular Multienzyme Complexes Quantum Theory Rats |
| spellingShingle |
Amidation of peptides Copper-containing proteins Peptidylglycine α-hydroxylating monooxygenase Peroxide copper H copper ion copper M cupric ion cuprous ion glycine hydrogen peroxide oxygen derivative oxygenase peptidylglycine alpha hydroxylating monooxygenase peroxide unclassified drug amidation animal cell article carboxy terminal sequence catalysis complex formation controlled study crystal structure energy enzyme active site enzyme binding enzyme metabolism female hamster hydroxylation isomer ligand binding molecular interaction molecular mechanics molecular model nonhuman priority journal quantum mechanics structure activity relation structure analysis X ray diffraction Animals Catalytic Domain CHO Cells Computer Simulation Coordination Complexes Copper Cricetinae Crystallography, X-Ray Hydrogen Bonding Hydrogen Peroxide Mixed Function Oxygenases Models, Molecular Multienzyme Complexes Quantum Theory Rats Coordination of peroxide to the CuM center of peptidylglycine α-hydroxylating monooxygenase (PHM): Structural and computational study |
| topic_facet |
Amidation of peptides Copper-containing proteins Peptidylglycine α-hydroxylating monooxygenase Peroxide copper H copper ion copper M cupric ion cuprous ion glycine hydrogen peroxide oxygen derivative oxygenase peptidylglycine alpha hydroxylating monooxygenase peroxide unclassified drug amidation animal cell article carboxy terminal sequence catalysis complex formation controlled study crystal structure energy enzyme active site enzyme binding enzyme metabolism female hamster hydroxylation isomer ligand binding molecular interaction molecular mechanics molecular model nonhuman priority journal quantum mechanics structure activity relation structure analysis X ray diffraction Animals Catalytic Domain CHO Cells Computer Simulation Coordination Complexes Copper Cricetinae Crystallography, X-Ray Hydrogen Bonding Hydrogen Peroxide Mixed Function Oxygenases Models, Molecular Multienzyme Complexes Quantum Theory Rats |
| description |
Many bioactive peptides, such as hormones and neuropeptides, require amidation at the C terminus for their full biological activity. Peptidylglycine α-hydroxylating monooxygenase (PHM) performs the first step of the amidation reaction - the hydroxylation of peptidylglycine substrates at the Cα position of the terminal glycine. The hydroxylation reaction is copper- and O2-dependent and requires 2 equiv of exogenous reductant. The proposed mechanism suggests that O2 is reduced by two electrons, each provided by one of two nonequivalent copper sites in PHM (CuH and CuM). The characteristics of the reduced oxygen species in the PHM reaction and the identity of the reactive intermediate remain uncertain. To further investigate the nature of the key intermediates in the PHM cycle, we determined the structure of the oxidized form of PHM complexed with hydrogen peroxide. In this 1.98-Å-resolution structure (hydro)peroxide binds solely to CuM in a slightly asymmetric side-on mode. The O-O interatomic distance of the copper-bound ligand is 1.5 Å, characteristic of peroxide/hydroperoxide species, and the Cu-O distances are 2.0 and 2.1 Å. Density functional theory calculations using the first coordination sphere of the CuM active site as a model system show that the computed energies of the side-on L3CuM(II)-O2 2- species and its isomeric, end-on structure L3CuM(I)-O 2 ·- are similar, suggesting that both these intermediates are significantly populated within the protein environment. This observation has important mechanistic implications. The geometry of the observed side-on coordinated peroxide ligand in L3CuM(II)O 2 2- is in good agreement with the results of a hybrid quantum mechanical-molecular mechanical optimization of this species. © 2012 SBIC. |
| title |
Coordination of peroxide to the CuM center of peptidylglycine α-hydroxylating monooxygenase (PHM): Structural and computational study |
| title_short |
Coordination of peroxide to the CuM center of peptidylglycine α-hydroxylating monooxygenase (PHM): Structural and computational study |
| title_full |
Coordination of peroxide to the CuM center of peptidylglycine α-hydroxylating monooxygenase (PHM): Structural and computational study |
| title_fullStr |
Coordination of peroxide to the CuM center of peptidylglycine α-hydroxylating monooxygenase (PHM): Structural and computational study |
| title_full_unstemmed |
Coordination of peroxide to the CuM center of peptidylglycine α-hydroxylating monooxygenase (PHM): Structural and computational study |
| title_sort |
coordination of peroxide to the cum center of peptidylglycine α-hydroxylating monooxygenase (phm): structural and computational study |
| publishDate |
2013 |
| url |
https://bibliotecadigital.exactas.uba.ar/collection/paper/document/paper_09498257_v18_n2_p223_Rudzka http://hdl.handle.net/20.500.12110/paper_09498257_v18_n2_p223_Rudzka |
| _version_ |
1768542948600840192 |