Peptidylglycine α‐amidating monooxygenase as a therapeutic target or biomarker for human diseases

Merkler, David J.; Hawley, Aidan J.; Eipper, Betty A.; Mains, Richard E.

doi:10.1111/bph.15815

Cited by 8 publications

(11 citation statements)

References 169 publications

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“…Numerous neuronal and endocrine processes rely on amidated peptides, while, PAM is the only bifunctional enzyme for amidated peptide biosynthesis ( 39 ). Although MR and co-localization analysis support that PAM increases the risk of insomnia, there are currently no clinical reports or studies exploring the underlying mechanisms.…”

Section: Discussionmentioning

confidence: 99%

Identification of potential drug targets for insomnia by Mendelian randomization analysis based on plasma proteomics

Yang,

Shi,

et al. 2024

Front. Neurol.

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IntroductionInsomnia, a common clinical disorder, significantly impacts the physical and mental well-being of patients. Currently, available hypnotic medications are unsatisfactory due to adverse reactions and dependency, necessitating the identification of new drug targets for the treatment of insomnia.MethodsIn this study, we utilized 734 plasma proteins as genetic instruments obtained from genome-wide association studies to conduct a Mendelian randomization analysis, with insomnia as the outcome variable, to identify potential drug targets for insomnia. Additionally, we validated our results externally using other datasets. Sensitivity analyses entailed reverse Mendelian randomization analysis, Bayesian co-localization analysis, and phenotype scanning. Furthermore, we constructed a protein-protein interaction network to elucidate potential correlations between the identified proteins and existing targets.ResultsMendelian randomization analysis indicated that elevated levels of TGFBI (OR = 1.01; 95% CI, 1.01–1.02) and PAM ((OR = 1.01; 95% CI, 1.01–1.02) in plasma are associated with an increased risk of insomnia, with external validation supporting these findings. Moreover, there was no evidence of reverse causality for these two proteins. Co-localization analysis confirmed that PAM (coloc.abf-PPH4 = 0.823) shared the same variant with insomnia, further substantiating its potential role as a therapeutic target. There are interactive relationships between the potential proteins and existing targets of insomnia.ConclusionOverall, our findings suggested that elevated plasma levels of TGFBI and PAM are connected with an increased risk of insomnia and might be promising therapeutic targets, particularly PAM. However, further exploration is necessary to fully understand the underlying mechanisms involved.

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Section: Discussionmentioning

confidence: 99%

Identification of potential drug targets for insomnia by Mendelian randomization analysis based on plasma proteomics

Yang,

Shi,

et al. 2024

Front. Neurol.

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“…In the biomedical arena, dopamine β‐monooxygenase (DBM) catalyzes a critical step in catecholamine biosynthesis (Klinman, 2006), as does its insect homologue tyramine β‐monooxygenase (TBM) (Hess et al, 2010). Similarly, peptidylglycine α‐amidating monooxygenase (PAM) (Merkler et al, 2022; Prigge et al, 2000; Welch et al, 2022a) is a bifunctional enzyme that is the only known enzyme to convert neuropeptide hormones into their active C‐terminally amidated forms. Here we focus on new insights into the structure and mechanism of the hydroxylase domain (peptidylglycine monooxygenase, PHM) of the bifunctional PAM enzyme.…”

Section: Introductionmentioning

confidence: 99%

New structures reveal flexible dynamics between the subdomains of peptidylglycine monooxygenase. Implications for an open to closed mechanism

2023

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Peptidylglycine monooxygenase (PHM) is essential for the biosynthesis of many neuroendocrine peptides via a copper‐dependent hydroxylation of a glycine‐extended pro‐peptide. The “canonical” mechanism requires the transfer of two electrons from one mononuclear copper (CuH, H‐site) to a second mononuclear copper (CuM, M‐site) which is the site of oxygen binding and catalysis. In most crystal structures the copper centers are separated by 11 Å of disordered solvent, but recent work has established that a PHM variant H108A forms a closed conformer in the presence of citrate with a reduced Cu–Cu site separation of ~4 Å. Here we report three new PHM structures where the H and M sites are separated by a longer distance of ~14 Å. Variation in Cu–Cu distance is the result of a rotation of the M subdomain about a hinge point centered on the pro199‐leu200‐ile201 triad which forms the linker between subdomains. The energetic cost of domain dynamics is likely small enough to allow free rotation of the subdomains relative to each other, adding credence to recent suggestions that an open‐to‐closed transition to form a binuclear oxygen binding intermediate is an essential element of catalysis. This inference would explain many experimental observations that are inconsistent with the current canonical mechanism including substrate‐induced oxygen activation and isotope scrambling during the peroxide shunt.

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“…PAM is a monopeptide, comprising two functional subunits with distinct catalytic activities, a peptidylglycine alpha-hydroxylating monooxygenase (PHM) and a peptidyl-alpha-hydroxyglycine alpha-amidating lyase (PAL) (Fig. 1 , inset right) 3 , 4 . Both catalytic domains work sequentially in order to convert a peptide intermediate into its active alpha-amidated form.…”

Section: Introductionmentioning

confidence: 99%

“…PAM is indispensable for life and is the only known enzyme to catalyze the C-terminal α-amidation 4 . It was demonstrated, that PAM double knock-out in mice and fruits flies is lethal in the first week of gestation or early larval stage, respectively, leading to a complete depletion of amidated peptide hormones 14 , 15 .…”

Section: Introductionmentioning

confidence: 99%

Immunoassay-based quantification of full-length peptidylglycine alpha-amidating monooxygenase in human plasma

Ilina,

Kaufmann,

Melander

et al. 2023

Sci Rep

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A one-step sandwich chemiluminescence immunometric assay (LIA) was developed for the quantification of bifunctional peptidylglycine-α-amidating monooxygenase (PAM) in human plasma (PAM-LIA). PAM is responsible for the activation of more than half of known peptide hormones through C-terminal α-amidation. The assay employed antibodies targeting specific catalytic PAM-subunits, peptidylglycine alpha-hydroxylating monooxygenase (PHM) and peptidyl-alpha-hydroxyglycine alpha-amidating lyase (PAL), to ensure detection of full-length PAM. The PAM-LIA assay was calibrated with a human recombinant PAM enzyme and achieved a detection limit of 189 pg/mL and a quantification limit of 250 pg/mL. The assay demonstrated good inter-assay (6.7%) and intra-assay (2.2%) variabilities. It exhibited linearity when accessed by gradual dilution or random mixing of plasma samples. The accuracy of the PAM-LIA was determined to be 94.7% through spiking recovery experiments, and the signal recovery after substance interference was 94–96%. The analyte showed 96% stability after six freeze–thaw cycles. The assay showed strong correlation with matched EDTA and serum samples, as well as matched EDTA and Li-Heparin samples. Additionally, a high correlation was observed between α-amidating activity and PAM-LIA. Finally, the PAM-LIA assay was successfully applied to a sub-cohort of a Swedish population-based study, comprising 4850 individuals, confirming its suitability for routine high throughput screening.

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Peptidylglycine α‐amidating monooxygenase as a therapeutic target or biomarker for human diseases

Cited by 8 publications

References 169 publications

Identification of potential drug targets for insomnia by Mendelian randomization analysis based on plasma proteomics

Identification of potential drug targets for insomnia by Mendelian randomization analysis based on plasma proteomics

New structures reveal flexible dynamics between the subdomains of peptidylglycine monooxygenase. Implications for an open to closed mechanism

Immunoassay-based quantification of full-length peptidylglycine alpha-amidating monooxygenase in human plasma

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