Monoclonal antibodies for human and porcine histamine N-methyltransferase (HMT) facilitate protein expression and localization studies

Schwelberger, H. G.; Feurle, Johannes

doi:10.1007/s00011-016-0987-1

Cited by 4 publications

(12 citation statements)

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“…In the central nervous system (CNS), histamine is metabolized by HNMT but not DAO, which is not found in the CNS. HNMT is typically found inside epithelial cells, hematopoietic cells, neurons, and glia . Histamine uptake into these cells is required for its metabolism.…”

Section: Introductionmentioning

confidence: 99%

Histamine and Migraine

Yuan

Silberstein

2017

Headache

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Background Histamine is an ancient “tissue amine” preceding multicellular organisms. In the central nervous system (CNS), its fibers originate solely from the tuberomammillary nucleus and travel throughout the brain. It is mainly responsible for wakefulness, energy homeostasis, and memory consolidation. Recently, several studies suggest a potential role of histamine in migraine pathogenesis and management. Methods Narrative review of current literature regarding histamine and migraine. Results Histamine plays a crucial role in migraine pathogenesis: sustaining the neurogenic inflammation pathway. Interaction between mast cells (MC) and calcitonin‐gene related protein (CGRP) results in sensitization of trigeminal afferents and trigeminal ganglia (TG). Histamine binds with differing affinities to four different histaminergic G‐protein coupled receptors, activating protein kinases, or triggering calcium release with subsequent mode of actions. Histamine 1 receptor (H1R) and histamine 2 receptor (H2R) antagonists are frequently used for the treatment of allergy and gastric acid secretion, respectively, but their antagonism is probably ineffective for migraine. Histamine 3 receptor (H3R) and histamine 4 receptor (H4R) have a threefold higher affinity than H1R/H2R for histamine and are found almost exclusively on neurons and immune tissues, respectively. H3R acts as an autoreceptor or as a heteroreceptor, lowering the release of histamine and other neurotransmitters. This is a potential target for anti‐nociception and anti‐neurogenic inflammation. To date, several small clinical trials using low dose histamine or Nα‐methylhistamine have demonstrated migraine prophylactic efficacy, probably via H3R or other undetermined pathways. Conclusion The histamine system interacts with multiple regions in the CNS and may hypothetically modulate the migraine response. Low dose histamine may be a promising option for migraine prevention.

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Section: Introductionmentioning

confidence: 99%

Histamine and Migraine

Yuan

Silberstein

2017

Headache

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“…Antibody clones HYB372-04/-05/-06/-07/-08/-09 resulted from immunization with human HNMT and clones HYB373-02/-03 resulted from immunization with porcine HNMT but cross-reacted with human HNMT [10]. To analyse where on the human HNMT protein these eight antibodies bind, a series of C-terminal deletions of the 292 amino acid HNMT protein was constructed by recombinant DNA technology and the resulting polypeptides were expressed as GST fusions in bacteria (Fig.…”

Section: Resultsmentioning

confidence: 99%

“…To characterize the human histamine-inactivating enzymes, we recently produced and characterized a series of mouse monoclonal antibodies specific for DAO and HNMT, respectively [ 9 , 10 ]. These antibodies turned out to be invaluable tools for the study of the expression and cellular localization of the enzymes.…”

Section: Introductionmentioning

confidence: 99%

“…These antibodies turned out to be invaluable tools for the study of the expression and cellular localization of the enzymes. The eight monoclonal antibodies binding human HNMT exhibited considerable differences in their binding characteristics and species cross-reactivity [ 10 ]. Additionally, we wanted to know if our antibodies recognize and can be used to detect the enzyme variants that have been described resulting from single nucleotide polymorphisms (SNPs) of the HNMT gene and that might be relevant for diseases associated with impaired histamine inactivation [ 11 , 12 ].…”

Section: Introductionmentioning

confidence: 99%

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Mapping of the binding sites of human histamine N-methyltransferase (HNMT) monoclonal antibodies

Schwelberger

Feurle

2017

Inflamm. Res.

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ObjectiveRecently, we characterized mouse monoclonal antibodies that allow the specific and sensitive detection of human histamine N-methyltransferase (HNMT). To understand differences in binding characteristics and recognition of enzyme variants we mapped the antibody binding sites.MethodsFragments of human HNMT were expressed as glutathione S-transferase fusion proteins that were used for testing antibody binding on immunoblots. Combined information from species cross-reactivity, sequence comparison, protein structure, and binding site prediction software were used to localize the epitope recognized by each antibody.ResultsAll eight monoclonal HNMT antibodies bound to linear epitopes in the C-terminal domain of the 292 amino acid protein. Of the five antibodies cross-reacting with HNMT from other species, one bound region L182–T223, three region M224–E261, and one region L262–A292. All three antibodies recognising only human HNMT bound the C-terminal region L262–A292 that contains residues present only in the human protein.ConclusionsOur HNMT monoclonal antibodies bind in three different regions of the protein and those binding the same putative epitope exhibit similar binding characteristics and species cross-reactivity. Antibodies binding non-overlapping epitopes will facilitate analyses of all clinically relevant variants described for HNMT.

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“…In order to characterize the human histamine inactivating enzymes, we recently produced and characterized a series of mouse monoclonal antibodies specific for DAO and HNMT, respectively [ 12 , 13 ]. These antibodies turned out to be invaluable tools for the study of the expression and cellular localization of the enzymes.…”

Section: Introductionmentioning

confidence: 99%

Mapping of the binding sites of human diamine oxidase (DAO) monoclonal antibodies

Schwelberger

Feurle

2017

Inflamm. Res.

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ObjectiveRecently we characterized five mouse monoclonal antibodies that allow the specific and sensitive detection of human diamine oxidase (DAO). To understand differences in binding characteristics and recognition of enzyme variants, we mapped the antibody binding sites.MethodsFragments of human DAO were expressed as glutathione-S-transferase fusion proteins that were used for testing antibody binding on immunoblots. Combined information from species cross-reactivity, sequence comparison and binding site-prediction software were used to localize the epitope recognized by each antibody.ResultsAll five monoclonal DAO antibodies bound to linear epitopes between the N3 and enzymatic domains of the 732 amino acid protein. The binding sites could be mapped onto amino acid regions V262-E278 and P279-R288, respectively, which exhibit considerable sequence variation in mammals explaining the fact that the human DAO antibodies do not cross-react with DAO from other species. The antibodies efficiently bind only denatured human DAO but not the native protein.ConclusionsCharacterization of the binding sites of the DAO antibodies revealed that the antibodies bind two adjacent epitopes and exhibit similar binding characteristics and species cross-reactivity. As the epitopes do not overlap any of the amino acid substitutions described for clinically significant DAO gene polymorphisms, our antibodies will also be useful for analyses of the mutant DAO proteins.

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Monoclonal antibodies for human and porcine histamine N-methyltransferase (HMT) facilitate protein expression and localization studies

Cited by 4 publications

References 32 publications

Histamine and Migraine

Histamine and Migraine

Mapping of the binding sites of human histamine N-methyltransferase (HNMT) monoclonal antibodies

Mapping of the binding sites of human diamine oxidase (DAO) monoclonal antibodies

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