Mannosidase activity of EDEM1 and EDEM2 depends on an unfolded state of their glycoprotein substrates

Shenkman, Marina; Ron, Efrat; Yehuda, Rivka; Benyair, Ron; Khalaila, Isam; Lederkremer, Gerardo Z.

doi:10.1038/s42003-018-0174-8

Cited by 49 publications

(50 citation statements)

References 42 publications

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“…Furthermore, even though CNX and CRT share substrates, some proteins are exclusive clients of each one (Molinari et al 2004;Lamriben et al 2016). On the other hand, phylogenetic analyses suggest that mns1(=ER mannosidase I) and edem (= ERAD-degradation-enhancinga-mannosidase-like protein) are also paralogs (Banerjee et al 2007) and both display mannosidase activity; however, EDEM preferentially recognizes misfolded proteins (Shenkman et al 2018).…”

Section: Discussionmentioning

confidence: 99%

Origin and Evolution of Two Independently Duplicated Genes Encoding UDP- Glucose: Glycoprotein Glucosyltransferases inCaenorhabditisand Vertebrates

Caraballo

Buzzi

Modenutti

et al. 2020

G3 Genes|Genomes|Genetics

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UDP-glucose: glycoprotein glucosyltransferase (UGGT) is a protein that operates as the gatekeeper for the endoplasmic reticulum (ER) quality control mechanism of glycoprotein folding. It is known that vertebrates and Caenorhabditis genomes harbor two uggt gene copies that exhibit differences in their properties.Bayesian phylogenetic inference based on 195 UGGT and UGGT-like protein sequences of an ample spectrum of eukaryotic species showed that uggt genes went through independent duplications in Caenorhabditis and vertebrates. In both lineages, the catalytic domain of the duplicated genes was subjected to a strong purifying selective pressure, while the recognition domain was subjected to episodic positive diversifying selection. Selective relaxation in the recognition domain was more pronounced in Caenorhabditis uggt-b than in vertebrates uggt-2. Structural bioinformatics analysis revealed that Caenorhabditis UGGT-b protein lacks essential sequences proposed to be involved in the recognition of unfolded proteins. When we assayed glucosyltrasferase activity of a chimeric protein composed by Caenorhabditis uggt-b recognition domain fused to S. pombe catalytic domain expressed in yeast, no activity was detected.The present results support the conservation of the UGGT activity in the catalytic domain and a putative divergent function of the recognition domain for the UGGT2 protein in vertebrates, which would have gone through a specialization process. In Caenorhabditis, uggt-b evolved under different constraints compared to uggt-a which, by means of a putative neofunctionalization process, resulted in a non-redundant paralog. The non-canonical function of uggt-b in the worm lineage highlights the need to take precautions before generalizing gene functions in model organisms. KEYWORDS

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Section: Discussionmentioning

confidence: 99%

Origin and Evolution of Two Independently Duplicated Genes Encoding UDP- Glucose: Glycoprotein Glucosyltransferases inCaenorhabditisand Vertebrates

Caraballo

Buzzi

Modenutti

et al. 2020

G3 Genes|Genomes|Genetics

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“…Initially, it was thought that the transition of the N-linked glycans to Man 8 GlcNAc 2 sorted the protein for ERAD [110]. ER Man I/Man1B1 has been implicated in removing the B-branch terminal mannose and recognizing tertiary and quaternary structure [111,112]. The activity of ERManI/Man1B1 is enhanced in the presence of the oxidoreductants PDI or TXNDC11 [112].…”

Section: Carbohydrate-dependent Protein Quality Controlmentioning

confidence: 99%

“…ER Man I/Man1B1 has been implicated in removing the B-branch terminal mannose and recognizing tertiary and quaternary structure [111,112]. The activity of ERManI/Man1B1 is enhanced in the presence of the oxidoreductants PDI or TXNDC11 [112]. In S. cerevisiae, the ER Man I equivalent, Mnslp, removes the outermost mannose from branch B, creating an eight mannose residue glycan (termed M8B) [110,113].…”

Section: Carbohydrate-dependent Protein Quality Controlmentioning

confidence: 99%

“…The EDEM family of three (EDEM1-3) serves as homologues to Htm1p. While all three EDEM family members have now been shown to possess mannosidase activity in cells, as well as using purified proteins more recently [119][120][121][122]112], the precise glycans they act upon and generate remains uncertain. Following the yeast paradigm, ER ManI/ Man1B1 and the EDEMs work together to create Man7-5 glycans with exposed 1,6-linked mannose residues on the C-branch [121].…”

Section: Carbohydrate-dependent Protein Quality Controlmentioning

confidence: 99%

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Protein Quality Control in the Endoplasmic Reticulum

2019

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The site of protein folding and maturation for the majority of proteins that are secreted, localized to the plasma membrane or targeted to endomembrane compartments is the endoplasmic reticulum (ER). It is essential that proteins targeted to the ER are properly folded in order to carry out their function, as well as maintain protein homeostasis, as accumulation of misfolded proteins could lead to the formation of cytotoxic aggregates. Because protein folding is an error-prone process, the ER contains protein quality control networks that act to optimize proper folding and trafficking of client proteins. If a protein is unable to reach its native state, it is targeted for ER retention and subsequent degradation. The protein quality control networks of the ER that oversee this evaluation or interrogation process that decides the fate of maturing nascent chains is comprised of three general types of families: the classical chaperones, the carbohydrate-dependent system, and the thiol-dependent system. The cooperative action of these families promotes protein quality control and protein homeostasis in the ER. This review will describe the families of the ER protein quality control network and discuss the functions of individual members.

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“…The glycomic profiles of free oligosaccharides (fOS) derived from misfolded N-and O-linked glycoproteins and lipid-linked oligosaccharides are important molecular signatures in various biological processes and serve as a readout of functional properties such as glycosidase inhibition. fOS profiles are useful for a diverse range of structural and functional studies such as glycoprotein folding (Hirayama 2018;Shenkman et al 2018), infant gut bacterial colonisation (Karav et al 2019), lactation (Liu et al 2019), bacterial (Nothaft et al 2010) and viral infections (Miller et al 2018), cancer (Alonzi et al 2011, and degenerative diseases such as osteoarthritis (Homan et al 2019), congenital-defects of glycosylation (CDG) (Davids et al 2019), and lysosomal storage disorders (Huang et al 2018). Additionally, fOS accumulation has also been used to evaluate the cell-based inhibition of glycosidase enzymes by small molecules (Glawar et al 2012;Rawlings et al 2009).…”

Section: Introductionmentioning

confidence: 99%

Analysis of Free Oligosaccharides (fOS) from Wild-Type Saccharomyces cerevisiae (Baker's Yeast) using Two Different Extraction Methods

Jalaludin¹,

Sudin²,

Elangovan³

et al. 2020

JSM

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The glycomic profiles of free oligosaccharides (fOS) derived from misfolded N-and O-linked glycoproteins and lipidlinked oligosaccharides are important molecular signatures in various biological processes and serve as a readout of functional properties such as glycosidase inhibition. Several glycan extraction methods are available based on different sorbent chemistries that may influence the analytical profiles obtained. However, there is limited availability of studies comparing the effects of sorbent chemistries on glycan profiles. Therefore, in our study, the fOS profiles from wild-type Saccharomyces cerevisiae (Baker's yeast) extracted using two common methods namely mixed-bed ion-exchange (MBIE) [AG50W-X12 (H +) and AG2-X8 (Cl-)] and reversed-phase (C18) sorbents were compared using total carbohydrate (phenol sulfuric acid) and total protein (bicinchoninic acid, BCA) assays, thin-layer chromatography (TLC) and highperformance liquid chromatography-evaporative light scattering detector (HPLC-ELSD) analyses. MBIE extraction contained higher oligosaccharide and protein (0.26 mg/mL and 1.8 mg/mL) content than C18 extraction (0.11 mg/mL and 0.2 mg/mL). TLC analysis (butanol: ethanol: water = 6:3:1 and 5:4:1) showed the presence of fOS in both the MBIE and C18 extracts based on the detection of orcinol active (UV-inactive) spots. Similar peaks were present in the HPLC-ELSD chromatograms for both extractions methods with MBIE showing higher abundance. Glycan unit (GU) analysis of the dextran standard using HPLC-ELSD showed that the largest possible oligosaccharide structures detected were only di/trisaccharides. Based on all these results, MBIE extraction is a more suitable carbohydrate extraction technique compared to C18 extraction for subsequent profiling and functional studies of fOS.

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Mannosidase activity of EDEM1 and EDEM2 depends on an unfolded state of their glycoprotein substrates

Cited by 49 publications

References 42 publications

Origin and Evolution of Two Independently Duplicated Genes Encoding UDP- Glucose: Glycoprotein Glucosyltransferases inCaenorhabditisand Vertebrates

Origin and Evolution of Two Independently Duplicated Genes Encoding UDP- Glucose: Glycoprotein Glucosyltransferases inCaenorhabditisand Vertebrates

Protein Quality Control in the Endoplasmic Reticulum

Analysis of Free Oligosaccharides (fOS) from Wild-Type Saccharomyces cerevisiae (Baker's Yeast) using Two Different Extraction Methods

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