Synthesis of glycoprotein molecular probes for the analyses of protein quality control system

Totani, Kiichiro; Matsuo, Ichiro; Takatani, Maki; Arai, Midori A.; Hagihara, Shinya; Ito, Yukishige

doi:10.1023/b:glyc.0000043751.18027.c1

Cited by 13 publications

(8 citation statements)

References 20 publications

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“…For this purpose, carbonic anhydrase (CA) and dihydrofolate reductase (DHFR) were chosen as model proteins. Glycan chains were to be introduced by either cystein alkylation (for CA) or ligand-carbohydrate conjugation (for DHFR) (63). CA is readily available as a homogeneous protein and DHFR can be expressed using either a cell-free system or a microbial expression system.…”

Section: C-5 Studies Of the Synthesis Of Glycoproteinsmentioning

confidence: 99%

My Stroll in the Backyard of Carbohydrate Chemistry

Ito

2010

TIGG

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The fields of "Glycobiology" and "Glycotechnology" are gaining an increasing amount of attention. A major part of my research has been directed at the synthesis and functional analysis of glycoconjugate oligosaccharides of various origins. In this mini-review, I wish to summarize my research history in carbohydrate chemistry of the last 12 years. Representative subjects have been 1) synthetic studies on Asn-linked glycan chains, 2) synthesis of novel glycoprotein structures, 3) approaches to polymer-support synthesis of oligosaccharides, 4) development of new reactions for the synthesis of glycoprotein-related molecules, 5) studies on the synthesis of glycoproteins, 6) analyses of glycoprotein processing and protein quality control system, 7) synthesis of glycosyltransferase inhibitors, 8) synthesis of biologically active glycolipids, and 9) synthesis and biological activities on mycobacterial cell wall components.

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Section: C-5 Studies Of the Synthesis Of Glycoproteinsmentioning

confidence: 99%

My Stroll in the Backyard of Carbohydrate Chemistry

Ito

2010

TIGG

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“…In this context, the most straightforward approach would be chemical ligation utilizing the unique reactivity of cysteine (Cys) residue on protein (Scheme ) 25. For instance, G0‐1Man9‐8GlcNAc2 was introduced to carbonic anhydrase (CA) to give artificial glycoprotein 28A,E,F 14b. CA is a relatively small (28 kD) protein having a single Cys residue.…”

Section: Synthesis Of N‐linked Glycoprotein‐related Molecular Probesmentioning

confidence: 99%

“…In order to gain more precise insight into glycoprotein quality control, we comprehensively synthesized high‐mannose‐type glycans12 and their partial structures 13. In addition, we prepared artificial glycoproteins carrying structurally defined and homogeneous oligosaccharides 14. By using these molecular probes, we conducted systematic and quantitative analyses of oligosaccharide–protein interactions 15…”

Section: Introductionmentioning

confidence: 99%

Exploration of oligosaccharide‐protein interactions in glycoprotein quality control by synthetic approaches

Hagihara

Totani

Ito

2006

The Chemical Record

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High-mannose-type oligosaccharides, which are cotranslationally introduced to nascent polypeptides, play important roles in glycoprotein quality control. This process is highly complex, involving a number of lectins, chaperones, and glycan-processing enzymes. For example, calnexin and calreticulin (CRT) are molecular chaperones that recognize monoglucosylated forms of high-mannose-type glycans. UDP-glucose : glycoprotein glucosyltransferase (UGGT) only glucosylates high-mannose-type glycans attached to partially folded proteins. Fbs1 is a component of ubiquitin ligase that recognizes sugar chains. Although recent studies have clarified the properties of these proteins, most of them used oligosaccharides derived from natural sources, which contain structural heterogeneity. In order to gain a more precise understanding, we started our program to comprehensively synthesize high-mannose-type glycans associated with a protein quality control system. Additionally, investigation of artificial glycoproteins led us to the discovery of the first nonpeptidic substrate of UGGT. These synthetic oligosaccharide probes have allowed us to conduct quantitative evaluations of the activity and specificity of CRT, Fbs1, and UGGT.

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“…[20,21] In order to gain a clear understanding of the glycoprotein quality-control system, structurally defined high-mannose-type glycans would be valuable tools. As part of our effort toward the comprehensive synthesis of ER-related glycoprotein glycans, [22] we have previously achieved the first chemical synthesis of a monoglucosylated dodecasaccharide (G1M9), [23] the putative ligand for CNX/CRT. We have also reported the first synthesis of Man 8 GlcNAc 2 (M8)-the proposed ligand of EDEM/Mnlp1/Htm1p-together with its monoglucosylated homologue Glc 1 Man 8 GlcNAc 2 (G1M8).…”

Section: Introductionmentioning

confidence: 99%

Design and Synthesis of Oligosaccharides that Interfere with Glycoprotein Quality‐control systems

et al. 2005

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Calnexin (CNX) and its soluble homologue calreticulin (CRT) are lectin-like molecular chaperones that help newly synthesized glycoproteins to fold correctly in the rough endoplasmic reticulum (ER). To investigate the mechanism of glycoprotein-quality control, we have synthesized structurally defined high-mannose-type oligosaccharides related to this system. This paper describes the synthesis of the non-natural undecasaccharide 2 and heptasaccharide 16, designed as potential inhibitors of the ER quality-control system. Each possesses the key tetrasaccharide element (Glc1Man3) critical for the CNX/CRT binding, while lacking the pentamannosyl branch required for glucosidase II recognition. These oligosaccharides were evaluated for their ability to bind CRT by isothermal titration calorimetry (ITC). As expected, each of them had a significant affinity towards CRT. In addition, these compounds were shown to be resistant to glucosidase II digestion. Their activities in blocking the chaperone function of CRT were next measured by using malate dehydrogenase (MDH) as a substrate. Their inhibitory effects were shown to correlate well with their CRT-binding affinities, both being critically dependent upon the presence of the terminal glucose (Glc) residue.

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Synthesis of glycoprotein molecular probes for the analyses of protein quality control system

Cited by 13 publications

References 20 publications

My Stroll in the Backyard of Carbohydrate Chemistry

My Stroll in the Backyard of Carbohydrate Chemistry

Exploration of oligosaccharide‐protein interactions in glycoprotein quality control by synthetic approaches

Design and Synthesis of Oligosaccharides that Interfere with Glycoprotein Quality‐control systems

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