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Proximity labeling of Siglec-1 counter-receptors on activated regulatory T cells using soluble Siglec-1-HRP multimers

ABSTRACT: Siglec-1 is a macrophage lectin-like receptor that mediates sialic acid-dependent cellular interactions. It was shown previously to promote inflammation in autoimmune disease through suppressing the expansion of regulatory T cells (Tregs). We have investigated the molecular basis for Siglec-1 binding to these cells using in vitro-induced Tregs. A proximity labelling and proteomics strategy were used to identify glycoproteins expressed by activated Tregs that may function as Siglec-1 counter-receptors.

INSTRUMENT(S): Q Exactive HF

ORGANISM(S): Homo Sapiens (human) Armoracia Rusticana Mus Musculus (mouse)

TISSUE(S): Cell Suspension Culture, T Cell

SUBMITTER: Gang Wu

LAB HEAD: Paul R. Crocker

PROVIDER: PXD021693 | Pride | 2021-04-28

REPOSITORIES: Pride

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			Action	DRS
	DeamidationNN1.zip	Other
	GW-E1.raw	Raw
	GW-E2.raw	Raw
	GW-E3.raw	Raw
	GW-F1.raw	Raw

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Proximity labeling of Siglec-1 counter-receptors on activated regulatory T cells using Siglec-1-HRP multimers on nano-beads

Project description:Siglec-1 is a macrophage lectin-like receptor that mediates sialic acid-dependent cellular interactions. It was shown previously to promote inflammation in autoimmune disease through suppressing the expansion of regulatory T cells (Tregs). We have investigated the molecular basis for Siglec-1 binding to these cells using in vitro-induced Tregs. A proximity labelling and proteomics strategy were used to identify glycoproteins expressed by activated Tregs that may function as Siglec-1 counter-receptors.

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Identification of Siglec-E-ligand candidates on BCL-1 mouse B cell line by proximity labeling

Project description:Siglec-E is a member of Siglec family of glycan recognition proteins involved in the regulation of myeloid cell activities. Siglec-E preferentially recognizes disialic acid, a glycan epitope consisting of two sialic acids conjugated in alpha2-8 linkage (Zhang JQ et al., Eur J Immunol. 2004, 34:1175-84). BCL-1, a mouse B cell line, expresses disialic acid epitope (Wang SH et al., Glycobiology. 2013, 23:677-89). In this experiment, we attempted at identifying the ligands of Siglec-E on BCL-1 by applying the proximity labeling method we developed (Chang L et al., J Proteome Res. 2017, 16:3929-41).

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Identification of Siglec-7-ligand candidates on JVM-3 human B cell line by proximity labeling

Project description:Siglec-7 is a member of Siglec family of glycan recognition proteins involved in the regulation of natural killer (NK) cell activities. We found Siglec-7 ligand is highly expressed on JVM-3, a B cell chronic lymphocytic leukemia (B-CLL) cell line. In this experiment, we attempted at identifying the ligands of Siglec-7 on JVM-3 by applying the proximity labeling method we developed (Chang L et al., J Proteome Res. 2017, 16:3929-41).

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Structural insights into Siglec-15 reveal glycosylation dependency for its interaction with T cells through integrin CD11b

Project description:Sialic acid-binding Ig-like lectin 15 (Siglec-15) is emerging as an immune modulator and target for cancer immunotherapy. However, limited knowledge regarding its mechanism of action and structure, or its binding partners with T cells, restrains the development of drug candidates that could unleash its full therapeutic potential. Here, we determine the crystal structure of Siglec-15 and delineate its binding epitope by co-crystallization with an anti-Siglec-15 blocking antibody. Saturation transfer-difference nuclear magnetic resonance (STD-NMR) spectroscopy assisted with molecular dynamics simulations reveals the binding mode of Siglec-15 to (2,3)- and 2,6)- linked sialic acids and to the cancer-associated sialyl-Tn (STn) glycoform. Binding of Siglec-15 to T cells, which lack STn expression, depends on the presence of (2,3)- and 2,6)- linked sialoglycans. We identify the leukocyte integrin CD11b as a binding partner of Siglec-15 on human T cells. Collectively, these data provide an integrative understanding of the structural features of Siglec-15 and underscore glycosylation as an additional layer of control of T cell responses.

2023-07-20 | PXD042009 | Pride

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2017-09-29 | PXD006359 | Pride

Gene expression profiles of Siglec-14/THP-1 and Siglec-5/THP cell lines, with or without NTHi stimulation

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2021-04-28 | PXD022259 | Pride

The Inhibitory Receptor Siglec-8 Interacts with FcεRI and Globally Inhibits Intracellular Signaling in Primary Mast Cells Upon Activation

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Identification of proteins that interact with Siglec-6

Project description:Siglec-6 is a glycoprotein overexpressed on chronic lymphocytic leukemia (CLL) cell. We have shown that Siglec-6 promotes migration and adhesion of CLL cells towards CLL bone marrow stromal cells. To elucidate the mechanistic pathway involved in Siglec-6 dependent migration, we performed mass spectrometry proteomics analysis on MEC1-002 CLL cell line pulled down with a Siglec-6 targeted antibody, which revealed that Siglec-6 interacts with DOCK8, a guanine nucleotide exchange factor. Interaction of Siglec-6 with its ligand promotes DOCK8 dependent Cdc42 activation, WASP protein recruitment and F-actin polymerization. Therapeutically, Siglec-6 leukemic cells can be eliminated with a Siglec-6 targeted bispecific antibody in vitro and in vivo.

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