How presence of a signal peptide affects human galectins-1 and -4: Clues to explain common absence of a leader sequence among adhesion/growth-regulatory galectins

Kutzner, Tanja J; Higuero, Alonso M.; Süßmair, Martina; Kopitz, Jürgen; Hingar, Michael; Díez-Revuelta, Natalia; Caballero, Gabriel García; Kaltner, Herbert; Lindner, Ingo; Abad‐Rodríguez, José; Reusch, Dietmar; Gabius, Hans‐Joachim

doi:10.1016/j.bbagen.2019.129449

Cited by 17 publications

(15 citation statements)

References 72 publications

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“…As a perspective, the availability of such variants, on the level of the cDNA and the protein, will facilitate a series of lines of functionally oriented research, starting with interrogating Gal-3 CRD-sharing proteins for consequences concerning the routes of galectin export from cells. After all, all galectins lack a signal sequence so that they first function intracellularly and then leave the cell by a variety of pathways (Hughes 1999 ; Sato 2018 ; Kutzner et al 2020 ). Considering the multifunctionality that galectins have intra- and extracellularly, and this with specificity for the cell type (and its special status of activation of differentiation) (Kasai 1997 , 2018 ; García Caballero et al 2020a ), work on more than a few assay systems will have to be done to characterize the role of this aspect of galectin structure reliably.…”

Section: Discussionmentioning

confidence: 99%

Influence of protein (human galectin-3) design on aspects of lectin activity

Caballero

Beckwith

Шилова

et al. 2020

Histochem Cell Biol

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The concept of biomedical significance of the functional pairing between tissue lectins and their glycoconjugate counterreceptors has reached the mainstream of research on the flow of biological information. A major challenge now is to identify the principles of structure–activity relationships that underlie specificity of recognition and the ensuing post-binding processes. Toward this end, we focus on a distinct feature on the side of the lectin, i.e. its architecture to present the carbohydrate recognition domain (CRD). Working with a multifunctional human lectin, i.e. galectin-3, as model, its CRD is used in protein engineering to build variants with different modular assembly. Hereby, it becomes possible to compare activity features of the natural design, i.e. CRD attached to an N-terminal tail, with those of homo- and heterodimers and the tail-free protein. Thermodynamics of binding disaccharides proved full activity of all proteins at very similar affinity. The following glycan array testing revealed maintained preferential contact formation with N-acetyllactosamine oligomers and histo-blood group ABH epitopes irrespective of variant design. The study of carbohydrate-inhibitable binding of the test panel disclosed up to qualitative cell-type-dependent differences in sections of fixed murine epididymis and especially jejunum. By probing topological aspects of binding, the susceptibility to inhibition by a tetravalent glycocluster was markedly different for the wild-type vs the homodimeric variant proteins. The results teach the salient lesson that protein design matters: the type of CRD presentation can have a profound bearing on whether basically suited oligosaccharides, which for example tested positively in an array, will become binding partners in situ. When lectin-glycoconjugate aggregates (lattices) are formed, their structural organization will depend on this parameter. Further testing (ga)lectin variants will thus be instrumental (i) to define the full range of impact of altering protein assembly and (ii) to explain why certain types of design have been favored during the course of evolution, besides opening biomedical perspectives for potential applications of the novel galectin forms.

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Section: Discussionmentioning

confidence: 99%

Influence of protein (human galectin-3) design on aspects of lectin activity

Caballero

Beckwith

Шилова

et al. 2020

Histochem Cell Biol

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“…This molecular process may provide dynamic structural flexibility for Gal-7 to adapt to a high-affinity, and even cross-linking, state from the mixed-conformer form of the homodimer upon association with non-glycan counter-receptors, and the observed difference in growth inhibition of neuroblastoma cells as a sign for a more active surface lattice at higher concentrations. Structural conversion then may bring about a similar outcome, as the presence of an N-glycan does, for a galectin, if directed to enter the endoplasmic reticulum [ 17 ]. Since galectins are at least bifunctional, they can engage in binding more than one partner.…”

Section: Discussionmentioning

confidence: 99%

“…Beyond this role as 'readers' and 'interpreters' of glycan-based signals within the concept of the sugar code [9,10], galectins also target peptide motifs, among them distinct cytoplasmic and nuclear proteins [11,12]. The common absence of a signal peptide allows galectins to gain access to the cytoplasm, and transport mechanisms ensure routing to the nucleus or their non-classical secretion [13][14][15][16][17]. Indeed, immunohistochemical studies show such a distribution profile for Gal-7 in cells and tissues (for examples, please see [18][19][20]).…”

Section: Introductionmentioning

confidence: 99%

Pro4 prolyl peptide bond isomerization in human galectin-7 modulates the monomer-dimer equilibrum to affect function

Miller

Nesmelova

Daragan

et al. 2020

Biochemical Journal

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Human galectin-7 (Gal-7; also termed p53-induced gene 1 product) acts as a multifunctional effector by productive pairing with distinct glycoconjugates and protein counter-receptors in the cytoplasm and nucleus, as well as on the cell surface. Its structural analysis by NMR spectroscopy detected doubling of a set of particular resonances, an indicator of Gal-7 existing in two conformational states in slow exchange on the chemical shift time scale. Structural positioning of this set of amino acids around the P4 residue and loss of this phenomenon in the bioactive P4L mutant indicated cis-trans isomerization at this site. Respective resonance assignments confirmed our proposal of two Gal-7 conformers. Mapping hydrogen bonds and considering van der Waals interactions in molecular dynamics simulations revealed a structural difference for the N-terminal peptide, with the trans-state being more exposed to solvent and more mobile than the cis-state. Affinity for lactose or glycan-inhibitable neuroblastoma cell surface contact formation was not affected, because both conformers associated with an overall increase in order parameters (S2). At low µM concentrations, homodimer dissociation is more favored for the cis-state of the protein than its trans-state. These findings give direction to mapping binding sites for protein counter-receptors of Gal-7, such as Bcl-2, JNK1, p53 or Smad3, and to run functional assays at low concentration to test the hypothesis that this isomerization process provides a (patho)physiologically important molecular switch for Gal-7.

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“…Strongly supporting the network concept, more than a single CG is often present in the same region, graphically underscored by the two-color staining with antibody preparations against two CGs and the resulting signal overlap. Typical for vertebrate galectins that all lack a signal peptide sequence and are produced on free ribosomes (Kutzner et al, 2020;Sato, 2018;Wilson, Firth, Powell, & Harrison, 1989), they are often found in the cytoplasm, sometimes in nuclei. Respective transport mechanisms are documented especially for mammalian galectin-3 (Haudek et al, 2010).…”

Section: Discussionmentioning

confidence: 99%

“…Methodologically, this set-up bringing plant lectins with a narrow specificity and galectins with less stringent limitation among β-galactosides together can thus help to delineate in situ binding partners for galectins on the level of glycans. However, there are two caveats: (i) organic solvents extract glycosphingolipids that are binding partners such as ganglioside GM1 (Ledeen, Kopitz, Abad-Rodríguez, & Gabius, 2018;Schwarz & Futerman, 1997) and (ii) our twocolor illustrations of localizing CG immunohistochemically and labeled CG by fluorescence microscopy disclose separate signal distributions: pairing in situ will thus make any bound glycan invisible for (ga)lectins added as probe. These F I G U R E 9 Galectin histochemical profiling of mesonephros.…”

Section: Discussionmentioning

confidence: 99%

Glycobiology of developing chicken kidney: Profiling the galectin family and selected β‐galactosides

Manning

Caballero

Ludwig

et al. 2020

The Anatomical Record

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The concept of the sugar code interprets the cellular glycophenotype as a rich source of information read by glycan-lectin recognition in situ. This study's aim is the comprehensive characterization of galectin expression by immunohistochemistry during chicken nephrogenesis along with mapping binding sites by (ga)lectin histochemistry. Light and two-color fluorescence microscopy were used. First, six plant/fungal lectins that are specific for galectin-binding parts of N-and O-glycans were applied. The spatiotemporally regulated distributions of these glycans in meso-and metanephros equip cells with potential binding partners for the galectins. Complete galectin profiling from HH Stage 20 (about 70-72 hr) onward revealed cell-, galectin-, and stage-dependent expression patterns. Representatives of all three types of modular architecture of the galectin family are detectable, and overlaps of signal distribution in light and two-color fluorescence microscopy illustrate a possibility for functional cooperation among them. Performing systematic galectin histochemistry facilitated comparisons between staining profiles of plant lectins and galectins. They revealed several cases for differences so that tissue lectins appear to be selective among the β-galactosides. Notably, selectivity is also disclosed in intrafamily comparison. Thus, combining experimental series with plant and tissue lectins is a means to characterize target populations of glycans presented by cellular glycoconjugates for individual galectins. Our results document the presence and sophisticated level of elaboration among β-galactosides and among the members of the family of galectins during organogenesis, using chicken galectins and kidney as model. Thus, they provide a clear guideline for functional assays using supramolecular tools, cells, and organ cultures.

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How presence of a signal peptide affects human galectins-1 and -4: Clues to explain common absence of a leader sequence among adhesion/growth-regulatory galectins

Cited by 17 publications

References 72 publications

Influence of protein (human galectin-3) design on aspects of lectin activity

Influence of protein (human galectin-3) design on aspects of lectin activity

Pro4 prolyl peptide bond isomerization in human galectin-7 modulates the monomer-dimer equilibrum to affect function

Glycobiology of developing chicken kidney: Profiling the galectin family and selected β‐galactosides

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