Abstract
We previously identified several peptide sequences that mimicked the terminal sugars of complex glycans. Using plant lectins as analogs of lectin-type cell-surface receptors, a tetravalent form of a peptide with the sequence NPSHPLSG, designated svH1C, bound with high avidity to lectins specific for glycans with terminal 5-acetylneuraminic acid (Neu5Ac)-galactose (Gal)/N-acetylgalactosamine (GalNAc) sequences. In this report, we show by circular dichroism and NMR spectra that svH1C lacks an ordered structure and thus interacts with binding sites from a flexible conformation. The peptide binds with high avidity to several recombinant human siglec receptors that bind preferentially to Neu5Ac(α2,3)Gal, Neu5Ac(α2,6)GalNAc or Neu5Ac(α2,8)Neu5Ac ligands. In addition, the peptide bound the receptor NKG2D, which contains a lectin-like domain that binds Neu5Ac(α2,3)Gal. The peptide bound to these receptors with a KD in the range of 0.6 to 1 μM. Binding to these receptors was inhibited by the glycoprotein fetuin, which contains multiple glycans that terminate in Neu5Ac(α2,3)Gal or Neu5Ac(α2,6)Gal, and by sialyllactose. Binding of svH1C was not detected with CLEC9a, CLEC10a or DC-SIGN, which are lectin-type receptors specific for other sugars. Incubation of neuraminidase-treated human peripheral blood mononuclear cells with svH1C resulted in binding of the peptide to a subset of the CD14+ monocyte population. Tyrosine phosphorylation of siglecs decreased dramatically when peripheral blood mononuclear cells were treated with 100 nM svH1C. Subcutaneous, alternate-day injections of svH1C into mice induced several-fold increases in populations of several types of immune cells in the peritoneal cavity. These results support the conclusion that svH1C mimics Neu5Ac-containing sequences and interacts with cell-surface receptors with avidities sufficient to induce biological responses at low concentrations. The attenuation of inhibitory receptors suggests that svH1C has characteristics of a checkpoint inhibitor.
Highlights
An extensive number of lectin-type cell-surface receptors regulate activity of immune cells [1]
Other examples of C-type lectins that undergo endocytosis include DC-SIGN/ CD209, a mannose (Man)-binding receptor on dendritic cells and macrophages; MRC1/ CD206, a Man receptor on macrophages; Langerin/CD207, a high Man and galactose-6-sulfated oligosaccharide receptor on Langerhans cells [6,7]; and Dectin-1/CLEC7a, a β-glucan receptor on macrophages [1]. Another large family of glycan-specific receptors includes I-type lectins that belong to the immunoglobulin superfamily
We previously found fetuin effectively competed with svH1C for binding to plant lectins, and that digestion of fetuin with neuraminidase nearly eliminated its ability to inhibit binding of svH1C [29]
Summary
An extensive number of lectin-type cell-surface receptors regulate activity of immune cells [1]. Other examples of C-type lectins that undergo endocytosis include DC-SIGN/ CD209, a mannose (Man)-binding receptor on dendritic cells and macrophages; MRC1/ CD206, a Man receptor on macrophages; Langerin/CD207, a high Man and galactose-6-sulfated oligosaccharide receptor on Langerhans cells [6,7]; and Dectin-1/CLEC7a, a β-glucan receptor on macrophages [1]. Another large family of glycan-specific receptors includes I-type lectins that belong to the immunoglobulin superfamily. Siglec-14, -15 and -16 lack the inhibitory domain and function in conjunction with an activating adaptor protein, DAP12, which contains an immunoreceptor tyrosine-based activation motif (ITAM) [11,12,17,18]
Sign-in/Register to view highlights & summary Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a callDisclaimer: All third-party content on this website/platform is and will remain the property of their respective owners and is provided on "as is" basis without any warranties, express or implied. Use of third-party content does not indicate any affiliation, sponsorship with or endorsement by them. Any references to third-party content is to identify the corresponding services and shall be considered fair use under The CopyrightLaw.
