Imaging the glycome

Laughlin, Scott T.; Bertozzi, Carolyn R.

doi:10.1073/pnas.0811481106

Cited by 346 publications

(314 citation statements)

References 90 publications

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“…In future studies, recently developed nontoxic click labeling reagents (47) and an expanding set of sugar analogs compatible with click chemistry (18) could extend the observations reported here by enabling the fine-scale characterization of the intracellular trafficking, delivery kinetics, and apoplastic modification of RG-I and other cell-wall polysaccharides in living seedlings. The specificity of FucAl incorporation we observed suggests that structural variants of sugar analogs might be differentially incorporated by their cognate glycosyltransferases, providing specific probes for individual cell-wall polysaccharides.…”

Section: Discussionmentioning

confidence: 99%

“…These sugar analogs are modified with an azido or alkynyl group and can be covalently coupled by a copper-catalyzed [2+3] cycloaddition "click" reaction to a fluorophore possessing the opposite functional group. This reaction is bio-orthogonal, proceeds rapidly at room temperature, and has been used to image glycoconjugates in multiple biological systems (18,19), but to date has not been applied to plant cell walls.…”

mentioning

confidence: 99%

“…The development of sugar analogs compatible with click chemistry (18) presents a unique opportunity to extend the suite of small-molecule cell-wall imaging tools. These sugar analogs are modified with an azido or alkynyl group and can be covalently coupled by a copper-catalyzed [2+3] cycloaddition "click" reaction to a fluorophore possessing the opposite functional group.…”

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confidence: 99%

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Metabolic click-labeling with a fucose analog reveals pectin delivery, architecture, and dynamics in Arabidopsis cell walls

Anderson

Wallace

Somerville

2012

Proc. Natl. Acad. Sci. U.S.A.

138

133

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Polysaccharide-rich cell walls are a defining feature of plants that influence cell division and growth, but many details of cell-wall organization and dynamics are unknown because of a lack of suitable chemical probes. Metabolic labeling using sugar analogs compatible with click chemistry has the potential to provide new insights into cell-wall structure and dynamics. Using this approach, we found that an alkynylated fucose analog (FucAl) is metabolically incorporated into the cell walls of Arabidopsis thaliana roots and that a significant fraction of the incorporated FucAl is present in pectic rhamnogalacturonan-I (RG-I). Time-course experiments revealed that FucAl-containing RG-I first localizes in cell walls as uniformly distributed punctae that likely mark the sites of vesiclemediated delivery of new polysaccharides to growing cell walls. In addition, we found that the pattern of incorporated FucAl differs markedly along the developmental gradient of the root. Using pulse-chase experiments, we also discovered that the pectin network is reoriented in elongating root epidermal cells. These results reveal previously undescribed details of polysaccharide delivery, organization, and dynamics in cell walls.fluorescence | secretion | small molecule T he primary cell walls of plants consist of a complex polysaccharide-rich network containing cellulose, hemicellulose, pectin, and structural proteins. Cellulose, which is the primary load-bearing component of the wall, is hydrogen-bonded to hemicelluloses, and this composite is embedded in a pectin matrix (1, 2). The cell wall must simultaneously maintain the structural integrity of the plant cell by resisting the osmotic pressure necessary for turgor-mediated cell growth, and remain sufficiently dynamic to expand along with the growing cell. These requirements are satisfied by the controlled synthesis, deposition, and alteration of cell-wall components. Cellulose is synthesized at the plasma membrane (3), whereas hemicelluloses and pectins are synthesized intracellularly and secreted into the wall via an incompletely characterized vesicle-mediated trafficking pathway (2).In contrast to proteins, for which genetically encoded fluorescent tags have provided many insights into localization and function, detailed characterization of cell-wall polysaccharide structure and dynamics in intact plants has proven difficult. Chemical (4, 5) and biochemical (6-8) techniques can provide primary structural information for extracted cell-wall polymers. Imaging approaches, including transmission electron microscopy (9, 10), Raman microspectroscopy (11), and studies using a growing array of lectins, carbohydrate-binding modules, and carbohydrate-specific antibodies (12) have provided information regarding the location or abundance of cell-wall polysaccharides in various cell types (13). However, these imaging techniques suffer from limitations, including long sample-preparation times, fixation artifacts, lack of temporal information, lack of polymer specificity (12), and masking of a...

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Section: Discussionmentioning

confidence: 99%

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confidence: 99%

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confidence: 99%

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Metabolic click-labeling with a fucose analog reveals pectin delivery, architecture, and dynamics in Arabidopsis cell walls

Anderson

Wallace

Somerville

2012

Proc. Natl. Acad. Sci. U.S.A.

138

133

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“…PLZF is critical for the function of innate‐like T‐cells, including MAIT cells,85 γδ T‐cells93 and iNKT cells 94, 95. Furthermore, newly described murine innate T‐cells such as the PLZF+ ROR γ t+ natural Th17 cells96 and PLZF+ T‐CD4 T‐cells97 also express PLZF, and are highly responsive to cytokines.…”

Section: Functional Similarities Between Nk Cell and Cd8+ T‐cell Subsetsmentioning

confidence: 99%

Innate‐like CD8+ T‐cells and NK cells: converging functions and phenotypes

2018

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SummaryNew data in the worlds of both innate‐like CD8+ T‐cells and natural killer (NK) cells have, in parallel, clarified some of the phenotypes of these cells and also their associated functions. While these cells are typically viewed entirely separately, the emerging innate functions of T‐cells and, similarly, the adaptive functions of NK cells suggest that many behaviours can be considered in parallel. In this review we compare the innate functions of CD8+ T‐cells (especially mucosal‐associated invariant T‐cells) and those of NK cells, and how these relate to expression of phenotypic markers, especially CD161 and CD56.

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“…Designing a strategy for biomarker discovery, new principles and platforms for the analysis of relatively small amounts of numerous glycoproteins and accurate monitoring at the level of the glycoproteome are gradually being developed (80)(81)(82). Screening of different types of glycomolecules, selection of differentially expressed components, their enrichment and purification or identi fication are the most challenging parts of experimental and clinical glycoproteomics.…”

Section: The »Omics« Era: the Glycome As A Source Of New Biomarkersmentioning

confidence: 99%

Glycans as Biomarkers: Status and Perspectives

Janković¹

2011

Journal of Medical Biochemistry

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lanaca koji su kovalentno vezani za poli pep tidnu ki~mu. Dobro je poznat zna~aj prome na u glikozilaciji proteina za nastanak, razvoj i krajnji ishod razli~itih bolesti kod ljudi. Glikani se smatraju jedinstvenim strukturama za di jag nozu, i pra}enje toka bolesti. U »omics« eri, glikom, glika n ski analog proteoma i genoma, predstavlja mogu}i izvor no vih biomarkera. Kreiranje strategije za otkri}e biomarkera zahteva nove principe i platforme za analizu relativno malih koli~ina brojnih glikoproteina. O~ekuje se da glikomske teh no logije, koje su jo{ u razvoju, a koje obuhvataju razli~ite tipove masene spektrometrije i afinitivnih tehnika, rezultiraju novim analiti~kim procedurama u oblasti odre |i vanja bio markera. Najve}i izazovi za eksperimentalnu i kli ni~ku gliko pro teomiku su: pretraga razli~itih tipova gliko mo lekula, oda bir potencijalnih markera i njihova selekcija ili pre ~i{}a -vanje i identifikacija. Za ovo je neophodno razviti teh no logije koje }e omogu}iti visoku senzitivnost detekcije biomarkera kao i odgovaraju}u standardizaciju i validaciju novih metoda, kako bi se one mogle primeniti u laboratorijskom radu. Dalji razvoj na polju primenjene glikonauke zahteva integrisani sistemski pristup sa ciljem da se na pravi na~in iskoriste sve njene mo gu}nosti u dijagnostici.

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Imaging the glycome

Cited by 346 publications

References 90 publications

Metabolic click-labeling with a fucose analog reveals pectin delivery, architecture, and dynamics in Arabidopsis cell walls

Metabolic click-labeling with a fucose analog reveals pectin delivery, architecture, and dynamics in Arabidopsis cell walls

Innate‐like CD8+ T‐cells and NK cells: converging functions and phenotypes

Glycans as Biomarkers: Status and Perspectives

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