Re-Evaluation of Binding Properties of Recombinant Lymphocyte Receptors NKR-P1A and CD69 to Chemically Synthesized Glycans and Peptides

Rozbeský, Daniel; Krejzová, Jana; Křenek, Karel; Prchal, Jan; Hrabal, Richard; Kožíšek, Milan; Weignerová, Lenka; Fiore, Michele; Dumy, Pascal; Křen, Vladimı́r; Renaudet, Olivier

doi:10.3390/ijms15011271

Cited by 7 publications

(5 citation statements)

References 35 publications

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“…NK receptors comprise two structurally divergent classes: the families of immunoglobulin-like receptors and C-type lectin-like receptors (CTLR) 3 , 4 . CTLRs are encoded within the natural killer gene complex (NKC, human chromosome 12), and, unlike C-type lectins, CTLRs neither bind calcium ions nor engage carbohydrate ligands 5 , 6 . Instead, CTLRs are known to interact with protein ligands.…”

Section: Introductionmentioning

confidence: 99%

Structure of the human NK cell NKR-P1:LLT1 receptor:ligand complex reveals clustering in the immune synapse

et al. 2022

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Signaling by the human C-type lectin-like receptor, natural killer (NK) cell inhibitory receptor NKR-P1, has a critical role in many immune-related diseases and cancer. C-type lectin-like receptors have weak affinities to their ligands; therefore, setting up a comprehensive model of NKR-P1-LLT1 interactions that considers the natural state of the receptor on the cell surface is necessary to understand its functions. Here we report the crystal structures of the NKR-P1 and NKR-P1:LLT1 complexes, which provides evidence that NKR-P1 forms homodimers in an unexpected arrangement to enable LLT1 binding in two modes, bridging two LLT1 molecules. These interaction clusters are suggestive of an inhibitory immune synapse. By observing the formation of these clusters in solution using SEC-SAXS analysis, by dSTORM super-resolution microscopy on the cell surface, and by following their role in receptor signaling with freshly isolated NK cells, we show that only the ligation of both LLT1 binding interfaces leads to effective NKR-P1 inhibitory signaling. In summary, our findings collectively support a model of NKR-P1:LLT1 clustering, which allows the interacting proteins to overcome weak ligand-receptor affinity and to trigger signal transduction upon cellular contact in the immune synapse.

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Section: Introductionmentioning

confidence: 99%

Structure of the human NK cell NKR-P1:LLT1 receptor:ligand complex reveals clustering in the immune synapse

et al. 2022

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“…Though the same experimental conditions (buffer, temperature, calcium concentration, a positive and a negative control sample etc. ) were used as described before, no binding was detected [ 77 ]. Moreover, the interaction analysis using a neoglycoconjugate binding assay revealed no affinity between rat Nkrp1a or mouse Nkrp1c and the various neoglycoproteins such as N -acetylglucosamine 36 BSA.…”

Section: Saccharide Ligands Of Nkrp1—20 Years Of Controversymentioning

confidence: 99%

Nkrp1 Family, from Lectins to Protein Interacting Molecules

et al. 2015

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The C-type lectin-like receptors include the Nkrp1 protein family that regulates the activity of natural killer (NK) cells. Rat Nkrp1a was reported to bind monosaccharide moieties in a Ca2+-dependent manner in preference order of GalNac > GlcNAc >> Fuc >> Gal > Man. These findings established for rat Nkrp1a have been extrapolated to all additional Nkrp1 receptors and have been supported by numerous studies over the past two decades. However, since 1996 there has been controversy and another article showed lack of interactions with saccharides in 1999. Nevertheless, several high affinity saccharide ligands were synthesized in order to utilize their potential in antitumor therapy. Subsequently, protein ligands were introduced as specific binders for Nkrp1 proteins and three dimensional models of receptor/protein ligand interaction were derived from crystallographic data. Finally, for at least some members of the NK cell C-type lectin-like proteins, the “sweet story” was impaired by two reports in recent years. It has been shown that the rat Nkrp1a and CD69 do not bind saccharide ligands such as GlcNAc, GalNAc, chitotetraose and saccharide derivatives (GlcNAc-PAMAM) do not directly and specifically influence cytotoxic activity of NK cells as it was previously described.

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“…The principle that fold does not necessarily imply function is well established in the case of the C-type carbohydrate-recognition domains of animal lectins, which are a subset of the broader family designated C-type lectin-like domains that includes many members that lack sugar-binding activity [ 5 ], including some for which reports of sugar-binding activity have recently been called into question [ 6 ]. In many cases, other target ligands, such as lipoproteins or other proteins, are known but in other instances the functions of these domains remain to be established.…”

Section: Carbohydrate Recognition In Multiple Protein Fold Familiesmentioning

confidence: 99%

Convergent and divergent mechanisms of sugar recognition across kingdoms

Taylor

Drickamer

2014

Current Opinion in Structural Biology

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Re-Evaluation of Binding Properties of Recombinant Lymphocyte Receptors NKR-P1A and CD69 to Chemically Synthesized Glycans and Peptides

Cited by 7 publications

References 35 publications

Structure of the human NK cell NKR-P1:LLT1 receptor:ligand complex reveals clustering in the immune synapse

Structure of the human NK cell NKR-P1:LLT1 receptor:ligand complex reveals clustering in the immune synapse

Nkrp1 Family, from Lectins to Protein Interacting Molecules

Convergent and divergent mechanisms of sugar recognition across kingdoms

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