New insights into copper monooxygenases and peptide amidation: Structure, mechanism and function

Sean Taylor Prigge, R. E. Mains, B. A. Eipper, Mario L Amzel

Research output: Contribution to journalArticle

Abstract

Many bioactive peptides must be amidated at their carboxy terminus to exhibit full activity. Surprisingly, the amides are not generated by a transamidation reaction. Instead, the hormones are synthesized from glycine-extended intermediates that are transformed into active amidated hormones by oxidative cleavage of the glycine N-Cα bond. In higher organisms, this reaction is catalyzed by a single bifunctional enzyme, peptidylglycine α-amidating monooxygenase (PAM). The PAM gene encodes one polypeptide with two enzymes that catalyze the two sequential reactions required for amidation. Peptidylglycine α-hydroxylating monooxygenase (PHM; EC 1.14.17.3) catalyzes the stereospecific hydroxylation of the glycine α-carbon of all the peptidylglycine substrates. The second enzyme, peptidyl-α-hydroxyglycine α-amidating lyase (PAL; EC 4.3.2.5) generates α-amidated peptide product and glyoxylate. PHM contains two redox-active copper atoms that, after reduction by ascorbate, catalyze the reduction of molecular oxygen for the hydroxylation of glycine-extended substrates. The structure of the catalytic core of rat PHM at atomic resolution provides a framework for understanding the broad substrate specificity of PHM, identifying residues critical for PHM activity, and proposing mechanisms for the chemical and electron-transfer steps in catalysis. Since PHM is homologous in sequence and mechanism to dopamine β-monooxygenase (DBM; EC 1.14.17.1), the enzyme that converts dopamine to norepinephrine during catecholamine biosynthesis, these structural and mechanistic insights are extended to DBM.

Original languageEnglish (US)
Pages (from-to)1236-1259
Number of pages24
JournalCellular and Molecular Life Sciences
Volume57
Issue number8-9
StatePublished - 2000

Fingerprint

Mixed Function Oxygenases
Glycine
Copper
Hydroxylation
Peptides
peptidylamidoglycolate lyase
Enzymes
Dopamine
Substrates
Hormones
Dopamine beta-Hydroxylase
Lyases
Molecular oxygen
Biosynthesis
Sequence Homology
Substrate Specificity
Catalysis
Amides
Oxidation-Reduction
Catecholamines

Keywords

  • Amidation
  • Ascorbate
  • Copper
  • Dopamine β-monooxygenase
  • Electron transfer
  • Oxygen chemistry
  • Peptide hormones
  • Peptidylglycine α-amidating monooxygenase
  • Structure

ASJC Scopus subject areas

  • Biochemistry, Genetics and Molecular Biology(all)
  • Cell Biology

Cite this

New insights into copper monooxygenases and peptide amidation : Structure, mechanism and function. / Prigge, Sean Taylor; Mains, R. E.; Eipper, B. A.; Amzel, Mario L.

In: Cellular and Molecular Life Sciences, Vol. 57, No. 8-9, 2000, p. 1236-1259.

Research output: Contribution to journalArticle

@article{cf4d5a2ef00a4f21a9b1ad04eb3c7c98,
title = "New insights into copper monooxygenases and peptide amidation: Structure, mechanism and function",
abstract = "Many bioactive peptides must be amidated at their carboxy terminus to exhibit full activity. Surprisingly, the amides are not generated by a transamidation reaction. Instead, the hormones are synthesized from glycine-extended intermediates that are transformed into active amidated hormones by oxidative cleavage of the glycine N-Cα bond. In higher organisms, this reaction is catalyzed by a single bifunctional enzyme, peptidylglycine α-amidating monooxygenase (PAM). The PAM gene encodes one polypeptide with two enzymes that catalyze the two sequential reactions required for amidation. Peptidylglycine α-hydroxylating monooxygenase (PHM; EC 1.14.17.3) catalyzes the stereospecific hydroxylation of the glycine α-carbon of all the peptidylglycine substrates. The second enzyme, peptidyl-α-hydroxyglycine α-amidating lyase (PAL; EC 4.3.2.5) generates α-amidated peptide product and glyoxylate. PHM contains two redox-active copper atoms that, after reduction by ascorbate, catalyze the reduction of molecular oxygen for the hydroxylation of glycine-extended substrates. The structure of the catalytic core of rat PHM at atomic resolution provides a framework for understanding the broad substrate specificity of PHM, identifying residues critical for PHM activity, and proposing mechanisms for the chemical and electron-transfer steps in catalysis. Since PHM is homologous in sequence and mechanism to dopamine β-monooxygenase (DBM; EC 1.14.17.1), the enzyme that converts dopamine to norepinephrine during catecholamine biosynthesis, these structural and mechanistic insights are extended to DBM.",
keywords = "Amidation, Ascorbate, Copper, Dopamine β-monooxygenase, Electron transfer, Oxygen chemistry, Peptide hormones, Peptidylglycine α-amidating monooxygenase, Structure",
author = "Prigge, {Sean Taylor} and Mains, {R. E.} and Eipper, {B. A.} and Amzel, {Mario L}",
year = "2000",
language = "English (US)",
volume = "57",
pages = "1236--1259",
journal = "Cellular and Molecular Life Sciences",
issn = "1420-682X",
publisher = "Birkhauser Verlag Basel",
number = "8-9",

}

TY - JOUR

T1 - New insights into copper monooxygenases and peptide amidation

T2 - Structure, mechanism and function

AU - Prigge, Sean Taylor

AU - Mains, R. E.

AU - Eipper, B. A.

AU - Amzel, Mario L

PY - 2000

Y1 - 2000

N2 - Many bioactive peptides must be amidated at their carboxy terminus to exhibit full activity. Surprisingly, the amides are not generated by a transamidation reaction. Instead, the hormones are synthesized from glycine-extended intermediates that are transformed into active amidated hormones by oxidative cleavage of the glycine N-Cα bond. In higher organisms, this reaction is catalyzed by a single bifunctional enzyme, peptidylglycine α-amidating monooxygenase (PAM). The PAM gene encodes one polypeptide with two enzymes that catalyze the two sequential reactions required for amidation. Peptidylglycine α-hydroxylating monooxygenase (PHM; EC 1.14.17.3) catalyzes the stereospecific hydroxylation of the glycine α-carbon of all the peptidylglycine substrates. The second enzyme, peptidyl-α-hydroxyglycine α-amidating lyase (PAL; EC 4.3.2.5) generates α-amidated peptide product and glyoxylate. PHM contains two redox-active copper atoms that, after reduction by ascorbate, catalyze the reduction of molecular oxygen for the hydroxylation of glycine-extended substrates. The structure of the catalytic core of rat PHM at atomic resolution provides a framework for understanding the broad substrate specificity of PHM, identifying residues critical for PHM activity, and proposing mechanisms for the chemical and electron-transfer steps in catalysis. Since PHM is homologous in sequence and mechanism to dopamine β-monooxygenase (DBM; EC 1.14.17.1), the enzyme that converts dopamine to norepinephrine during catecholamine biosynthesis, these structural and mechanistic insights are extended to DBM.

AB - Many bioactive peptides must be amidated at their carboxy terminus to exhibit full activity. Surprisingly, the amides are not generated by a transamidation reaction. Instead, the hormones are synthesized from glycine-extended intermediates that are transformed into active amidated hormones by oxidative cleavage of the glycine N-Cα bond. In higher organisms, this reaction is catalyzed by a single bifunctional enzyme, peptidylglycine α-amidating monooxygenase (PAM). The PAM gene encodes one polypeptide with two enzymes that catalyze the two sequential reactions required for amidation. Peptidylglycine α-hydroxylating monooxygenase (PHM; EC 1.14.17.3) catalyzes the stereospecific hydroxylation of the glycine α-carbon of all the peptidylglycine substrates. The second enzyme, peptidyl-α-hydroxyglycine α-amidating lyase (PAL; EC 4.3.2.5) generates α-amidated peptide product and glyoxylate. PHM contains two redox-active copper atoms that, after reduction by ascorbate, catalyze the reduction of molecular oxygen for the hydroxylation of glycine-extended substrates. The structure of the catalytic core of rat PHM at atomic resolution provides a framework for understanding the broad substrate specificity of PHM, identifying residues critical for PHM activity, and proposing mechanisms for the chemical and electron-transfer steps in catalysis. Since PHM is homologous in sequence and mechanism to dopamine β-monooxygenase (DBM; EC 1.14.17.1), the enzyme that converts dopamine to norepinephrine during catecholamine biosynthesis, these structural and mechanistic insights are extended to DBM.

KW - Amidation

KW - Ascorbate

KW - Copper

KW - Dopamine β-monooxygenase

KW - Electron transfer

KW - Oxygen chemistry

KW - Peptide hormones

KW - Peptidylglycine α-amidating monooxygenase

KW - Structure

UR - http://www.scopus.com/inward/record.url?scp=0000821179&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0000821179&partnerID=8YFLogxK

M3 - Article

C2 - 11028916

AN - SCOPUS:0000821179

VL - 57

SP - 1236

EP - 1259

JO - Cellular and Molecular Life Sciences

JF - Cellular and Molecular Life Sciences

SN - 1420-682X

IS - 8-9

ER -