Characterization of a New Glycosynthase Cloned by Using Chemical Complementation

Tao, Hai‐Yan; Peralta‐Yahya, Pamela; Decatur, John; Cornish, Virginia W.

doi:10.1002/cbic.200700545

Cited by 16 publications

(9 citation statements)

References 44 publications

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“…The enzyme-catalyzed glycosylation reactions are not accompanied by the release of a chromophore, so novel screens were needed. The approaches that have been devised fall into three categories: a yeast three-hybrid chemical complementation assay (31), an assay based on pH changes (32), and a fluorescence-activating cell sorting (FACS) assay (33). In all three approaches, glycosynthase activity is evaluated in whole cells; therefore, protein isolation is not required.…”

Section: Figurementioning

confidence: 99%

Chemical Approaches to Glycobiology

Kiessling

Splain

2010

Annu. Rev. Biochem.

208

176

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Glycans are ubiquitous components of all organisms. Efforts to elucidate glycan function and to understand how they are assembled and disassembled can reap benefits in fields ranging from bioenergy to human medicine. Significant advances in our knowledge of glycan biosynthesis and function are emerging, and chemical biology approaches are accelerating the pace of discovery. Novel strategies for assembling oligosaccharides, glycoproteins, and other glycoconjugates are providing access to critical materials for interrogating glycan function. Chemoselective reactions that facilitate the synthesis of glycan-substituted imaging agents, arrays, and materials are yielding compounds to interrogate and perturb glycan function and dysfunction. To complement these advances, small molecules are being generated that inhibit key glycan-binding proteins or biosynthetic enzymes. These examples illustrate how chemical glycobiology is providing new insight into the functional roles of glycans and new opportunities to interfere with or exploit these roles.

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Section: Figurementioning

confidence: 99%

Chemical Approaches to Glycobiology

Kiessling

Splain

2010

Annu. Rev. Biochem.

208

176

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“…In the presence of an appropriate glycosynthase, transcriptional activation of a LEU2 gene occurred, as evidenced by increased survival on leucine dropout medium. This system has been successfully applied to identify a novel glycosynthase (51). …”

Section: Techniques Requiring Genetic Manipulationmentioning

confidence: 99%

Design and Applications of Bifunctional Small Molecules: Why Two Heads Are Better Than One

2008

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Induction of protein-protein interactions is a daunting challenge, but recent studies show promise for small molecules that specifically bring two or more protein molecules together for enhanced or novel biological effect. The first such bifunctional molecules were the rapamycin- and FK506-based “Chemical Inducers of Dimerization”, but the field has since expanded with new molecules and new applications in chemical genetics and cell biology. Examples include coumermycin-mediated gyrase B dimerization, proteolysis targeting chimeric molecules (PROTACS), drug hybrids, and strategies for exploiting multivalency in toxin binding and antibody recruitment. This review discusses these and other advances in the design and use of bifunctional small molecules, and potential strategies for future systems.

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“…Also progress in methods for identification and characterization of glycosynthases from mutant libraries will be critical to produce enzymes with new acceptor or donor scopes. Many advances have already been made such as biological and biochemical assays utilizing chemical complementation, fluorescence and photometric methods [ 50 , 88 , 89 , 90 ]. Subsequent analysis of structural and mechanistic details will help to identify structural requirements, which can be transferred to new glycosynthases by genetic engineering and rational design [ 28 , 91 ].…”

Section: Discussionmentioning

confidence: 99%

Synthesis of Glycosides by Glycosynthases

Hayes

Pietruszka

2017

Molecules

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The many advances in glycoscience have more and more brought to light the crucial role of glycosides and glycoconjugates in biological processes. Their major influence on the functionality and stability of peptides, cell recognition, health and immunity and many other processes throughout biology has increased the demand for simple synthetic methods allowing the defined syntheses of target glycosides. Additional interest in glycoside synthesis has arisen with the prospect of producing sustainable materials from these abundant polymers. Enzymatic synthesis has proven itself to be a promising alternative to the laborious chemical synthesis of glycosides by avoiding the necessity of numerous protecting group strategies. Among the biocatalytic strategies, glycosynthases, genetically engineered glycosidases void of hydrolytic activity, have gained much interest in recent years, enabling not only the selective synthesis of small glycosides and glycoconjugates, but also the production of highly functionalized polysaccharides. This review provides a detailed overview over the glycosylation possibilities of the variety of glycosynthases produced until now, focusing on the transfer of the most common glucosyl-, galactosyl-, xylosyl-, mannosyl-, fucosyl-residues and of whole glycan blocks by the different glycosynthase enzyme variants.

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Characterization of a New Glycosynthase Cloned by Using Chemical Complementation

Cited by 16 publications

References 44 publications

Chemical Approaches to Glycobiology

Chemical Approaches to Glycobiology

Design and Applications of Bifunctional Small Molecules: Why Two Heads Are Better Than One

Synthesis of Glycosides by Glycosynthases

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