A role for N-glycosylation in active adenosine deaminase 2 production

Ito, Moeko; Maejima, Yuko; Nishimura, Kazuki; Nakae, Yusei; Ono, Ayami; Iwaki-Egawa, Sachiko

doi:10.1016/j.bbagen.2022.130237

Cited by 1 publication

(11 citation statements)

References 24 publications

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“…We observed that intracellular dimer formation largely correlated with impairment of ADA2 secretion although even pathogenic variants with residual secretion, p.G47R, p.N127Q and p.P344L, showed increased dimer formation compared to WT. Inhibition of N-glycosylation -but not of the secretory pathwayled to aggregate formation of WT ADA2 (Figure 5B), confirming the role of N-glycosylation in ensuring the structural integrity of ADA2 (Ito et al, 2022). We also detected intracellular dimers in DADA2 HMDM although the signal was weaker due to the strongly reduced protein expression of the pathogenic variants (Figure 5C).…”

Section: Pathogenic Ada2 Variants Form Intracellular Dimerssupporting

confidence: 64%

Section: Discussionmentioning

confidence: 99%

“…The importance of N-glycosylation in trafficking and function of ADA2 has previously been established (11,12). Yet, the behavior of mutant ADA2 on the cellular level has to date only been examined in overexpression systems of cells that do not express the protein endogenously (12,13,19). Importantly, expression of glycosyltransferases and glycosidases and therefore glycosylation patterns of individual proteins show striking variation even between different immune cell populations (25).…”

Section: Discussionmentioning

confidence: 99%

“…Differences in the molecular weight of a protein can be caused by posttranslational protein modifications and N-glycosylation of ADA2 has previously been confirmed (10,12,17). Therefore, we initially hypothesized that the LMW-form of ADA2 unique to cells expressing WT ADA2 corresponded to glycan-free ADA2.…”

Section: Human Monocyte-derived Macrophages Intracellularly Express A...mentioning

confidence: 98%

“…monocyte-derived macrophages intracellularly express a hypoglycosylated low-molecular-weight form of ADA2 that is absent in DADA2 macrophages Differences in the molecular weight of a protein can be caused by posttranslational protein modifications and N-glycosylation of ADA2 has previously been confirmed (Zavialov et al, 2010;Greiner-Tollersrud et al, 2020;Ito et al, 2022). Therefore, we initially hypothesized that the LMWform of ADA2 present only in cells expressing WT ADA2 corresponded to glycan-free ADA2.…”

Section: Humanmentioning

confidence: 99%

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Human ADA2 deficiency is characterized by the absence of an intracellular hypoglycosylated form of adenosine deaminase 2

Ehlers,

Hombrouck,

Wouters

et al. 2023

Preprint

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Pathogenic variants inADA2underlie human deficiency of adenosine deaminase 2 (DADA2) and lead to impaired ADA2 protein secretion and reduced deaminase activity. Yet, the mechanisms driving the disease on a cellular level are poorly understood. Here, we report thatin vitrodifferentiation of macrophages from DADA2 CD14+ monocytes partially restores protein expression of mutant ADA2. Healthy control macrophages express a high-molecular-weight (HMW) secreted form of ADA2 and a hypoglycosylated low-molecular-weight (LMW) intracellular protein. LMW-ADA2 is absent in DADA2 macrophages, suggesting differential glycosylation of mutant ADA2. In HEK293T cells transfected with pathogenic ADA2 variants, HMW-ADA2 accumulates intracellularly. Interactome analysis of the pathogenic variant p.R169Q revealed increased binding to proteins of the protein folding machinery compared with wild-type ADA2. Pathogenic ADA2 variants form intracellular protein aggregates and show increased degradation upon inhibition of the secretory pathway. In conclusion, we show aberrant N-glycosylation of ADA2 expressed in DADA2 macrophages for the first time and establish the absence of LMW-ADA2 as a characteristic of these ADA2-deficient cells.Graphical abstractFigure 1:SummaryThis study identifies a previously undescribed hypoglycosylated intracellular form of ADA2 in human monocyte-derived macrophages. Absence of this low-molecular-weight form is a key feature of human ADA2 deficiency (DADA2), suggesting a role for intracellular ADA2 in the pathogenesis of the disease.

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Section: Pathogenic Ada2 Variants Form Intracellular Dimerssupporting

confidence: 64%

Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Section: Human Monocyte-derived Macrophages Intracellularly Express A...mentioning

confidence: 98%

Section: Humanmentioning

confidence: 99%

See 3 more Smart Citations

Human ADA2 deficiency is characterized by the absence of an intracellular hypoglycosylated form of adenosine deaminase 2

Ehlers,

Hombrouck,

Wouters

et al. 2023

Preprint

View full text Add to dashboard Cite

show abstract

A role for N-glycosylation in active adenosine deaminase 2 production

Cited by 1 publication

References 24 publications

Human ADA2 deficiency is characterized by the absence of an intracellular hypoglycosylated form of adenosine deaminase 2

Human ADA2 deficiency is characterized by the absence of an intracellular hypoglycosylated form of adenosine deaminase 2

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