Characterization of the LM5 pectic galactan epitope with synthetic analogues of β-1,4-d-galactotetraose

Andersen, Mathias Christian Franch; Boos, Irene; Marcus, Susan E.; Kračun, Stjepan Krešimir; Rydahl, Maja Gro; Willats, William G. T.; Knox, John; Clausen, Mads Hartvig

doi:10.1016/j.carres.2016.10.012

Cited by 28 publications

(27 citation statements)

References 24 publications

(31 reference statements)

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“…The binding of the 1,4-galactan-specific MAb LM5 is shown for comparison and its recognition capabilities are restricted to the set of linear oligo-b-1,4-galactosides and not those with substitutions (Fig. 3), as recently reported (Andersen et al, 2016b), indicating the complementary recognition of branched and linear galactan chains by LM26 and LM5, respectively.…”

Section: Monoclonal Antibody Lm26 Binds Specifically To Phloem Sieve supporting

confidence: 65%

“…Welldefined, single-compound oligosaccharides provide the highest resolution possible when characterizing protein-carbohydrate interactions, provided that the binding interactions are sufficiently strong to be detected. This is especially true when a range of structurally related but distinct glycans are available (for examples, see Clausen et al, 2003;Andersen et al, 2016aAndersen et al, , 2016bSchmidt et al, 2015). In the case of LM26 discussed here, it would have been close to impossible to identify the epitope without access to the appropriate and defined linear and branched galactans.…”

Section: Discussion Synthetic Glycan Arrays As a Discovery Platformmentioning

confidence: 97%

“…However, the micromechanical basis of galactanrich RG-I contributions to cell wall firmness are not known. It has recently been determined that the LM5 epitope encompasses terminal galactosyl residues at the nonreducing end of 1,4-galactan (Andersen et al, 2016b) and this epitope has been specifically located at inner cell wall regions adjacent to plasma membranes (McCartney et al, 2000;Lee et al, 2013)-a location observed for the LM26 branched galactan epitope (Figs. 2 and 5).…”

Section: Functional Role Of the Modulation Of 14-galactan Rg-i Strucmentioning

confidence: 96%

“…LM26 was used in the form of unpurified hybridoma cell culture supernatant for all analyses except for immunogold TEM. Other MAbs used in this study include LM5 to pectic 1,4-galactan (Jones et al, 1997;Andersen et al, 2016b) and JIM7 (Clausen et al, 2003) and LM19 (Verhertbruggen et al, 2009) to pectic homogalacturonan; and xyloglucan MAb LM25 .…”

Section: Monoclonal Antibodiesmentioning

confidence: 99%

“…3) has been described previously Andersen et al, 2016a). Carbohydrate microarray screening was performed as described previously (Andersen et al, 2016a(Andersen et al, , 2016b. Briefly, deprotected oligosaccharides were functionalized with a heterobifunctional linker 2-(N-methylaminooxy)-1-ethanamine, purified from the excess linker and printed onto NHS-activated Slide-H microarrays from Schott using the ArrayJet Sprint microarray printer.…”

Section: Synthetic Glycan/galactan Microarraysmentioning

confidence: 99%

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Branched Pectic Galactan in Phloem-Sieve-Element Cell Walls: Implications for Cell Mechanics

et al. 2017

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A major question in plant biology concerns the specification and functional differentiation of cell types. This is in the context of constraints imposed by networks of cell walls that both adhere cells and contribute to the form and function of developing organs. Here, we report the identification of a glycan epitope that is specific to phloem sieve element cell walls in several systems. A monoclonal antibody, designated LM26, binds to the cell wall of phloem sieve elements in stems of Arabidopsis (Arabidopsis thaliana), Miscanthus x giganteus, and notably sugar beet (Beta vulgaris) roots where phloem identification is an important factor for the study of phloem unloading of Suc. Using microarrays of synthetic oligosaccharides, the LM26 epitope has been identified as a b-1,6-galactosyl substitution of b-1,4-galactan requiring more than three backbone residues for optimized recognition. This branched galactan structure has previously been identified in garlic (Allium sativum) bulbs in which the LM26 epitope is widespread throughout most cell walls including those of phloem cells. Garlic bulb cell wall material has been used to confirm the association of the LM26 epitope with cell wall pectic rhamnogalacturonan-I polysaccharides. In the phloem tissues of grass stems, the LM26 epitope has a complementary pattern to that of the LM5 linear b-1,4-galactan epitope, which is detected only in companion cell walls. Mechanical probing of transverse sections of M. x giganteus stems and leaves by atomic force microscopy indicates that phloem sieve element cell walls have a lower indentation modulus (indicative of higher elasticity) than companion cell walls.

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Section: Monoclonal Antibody Lm26 Binds Specifically To Phloem Sieve supporting

confidence: 65%