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Human diamine oxidase cellular binding and internalization in vitro and rapid clearance in vivo are not mediated by N-glycans but by heparan sulfate proteoglycan interactions
- Source :
- Glycobiology. 31(4)
- Publication Year :
- 2020
-
Abstract
- Human diamine oxidase (hDAO) rapidly inactivates histamine by deamination. No pharmacokinetic data are available to better understand its potential as a new therapeutic modality for diseases with excess local and systemic histamine, like anaphylaxis, urticaria or mastocytosis. After intravenous administration of recombinant hDAO to rats and mice, more than 90% of the dose disappeared from the plasma pool within 10 min. Human DAO did not only bind to various endothelial and epithelial cell lines in vitro, but was also unexpectedly internalized and visible in granule-like structures. The uptake of rhDAO into cells was dependent on neither the asialoglycoprotein-receptor (ASGP-R) nor the mannose receptor (MR) recognizing terminal galactose or mannose residues, respectively. Competition experiments with ASGP-R and MR ligands did not block internalization in vitro or rapid clearance in vivo. The lack of involvement of N-glycans was confirmed by testing various glycosylation mutants. High but not low molecular weight heparin strongly reduced the internalization of rhDAO in HepG2 cells and HUVECs. Human DAO was readily internalized by CHO-K1 cells, but not by the glycosaminoglycan- and heparan sulfate-deficient CHO cell lines pgsA-745 and pgsD-677, respectively. A docked heparin hexasaccharide interacted well with the predicted heparin binding site 568RFKRKLPK575. These results strongly imply that rhDAO clearance in vivo and cellular uptake in vitro is independent of N-glycan interactions with the classical clearance receptors ASGP-R and MR, but is mediated by binding to heparan sulfate proteoglycans followed by internalization via an unknown receptor.
- Subjects :
- Glycosylation
media_common.quotation_subject
Mannose
CHO Cells
Biochemistry
03 medical and health sciences
chemistry.chemical_compound
Mice
0302 clinical medicine
In vivo
Cricetinae
medicine
Animals
Humans
Internalization
030304 developmental biology
media_common
Glycosaminoglycans
0303 health sciences
Heparin
In vitro
Cell biology
Rats
chemistry
Amine Oxidase (Copper-Containing)
Heparitin Sulfate
Diamine oxidase
030217 neurology & neurosurgery
Mannose receptor
Heparan Sulfate Proteoglycans
medicine.drug
Subjects
Details
- ISSN :
- 14602423
- Volume :
- 31
- Issue :
- 4
- Database :
- OpenAIRE
- Journal :
- Glycobiology
- Accession number :
- edsair.doi.dedup.....e2c9f609b585d05f9d314f4f93dbab6f