tailieunhanh - Báo cáo khoa học: Frontal affinity chromatography analysis of constructs of DC-SIGN, DC-SIGNR and LSECtin extend evidence for affinity to agalactosylated N-glycans

Dendritic cell-specific intracellular adhesion molecule-3-grabbing noninte-grin (DC-SIGN) is a member of the C-type lectin family selectively expressed on immune-related cells. In the present study, we performed a systematic interaction analysis of DC-SIGN and its related receptors, DC-SIGN-related protein (DC-SIGNR) and liver and lymph node sinusoi-dal endothelial cell C-type lectin (LSECtin) using frontal affinity chroma-tography (FAC). | Frontal affinity chromatography analysis of constructs of DC-SIGN DC-SIGNR and LSECtin extend evidence for affinity to agalactosylated N-glycans Rikio Yabe Hiroaki Tateno and Jun Hirabayashi Research Center for MedicalGlycoscience National institute of Advanced Industrial science and Technology AIST Tsukuba Ibaraki Japan Keywords agalactosylated N-glycan C-type lectin DC-SIGN DC-SIGNR LSECtin Correspondence J. Hirabayashi Research Center for Medical Glycoscience NationalInstitute of Advanced IndustrialScience and Technology AIST Tsukuba Ibaraki 305-8568 Japan Fax 81 29 861 3125 Tel 81 29 861 3124 E-mail jun-hirabayashi@ Received 19 August 2009 revised 22 June 2010 accepted 27 July 2010 doi Dendritic cell-specific intracellular adhesion molecule-3-grabbing noninte-grin DC-SIGN is a member of the C-type lectin family selectively expressed on immune-related cells. In the present study we performed a systematic interaction analysis of DC-SIGN and its related receptors DC-SIGN-related protein DC-SIGNR and liver and lymph node sinusoidal endothelial cell C-type lectin LSECtin using frontal affinity chromatography FAC . Carbohydrate-recognition domains of the lectins expressed as Fc-fusion chimeras were immobilized to Protein A-Sepharose and subjected to quantitative FAC analysis using 157 pyridylaminated gly-cans. Both DC-SIGN-Fc and DC-SIGNR-Fc showed similar specificities for glycans containing terminal mannose and fucose but great difference in affinity under the given experimental conditions. By contrast LSECtin-Fc showed no affinity to these glycans. As a common feature the DC-SIGN-related lectin-Fc chimeras including LSECtin exhibited binding affinity to mono- and or bi-antennary agalactosylated N-glycans. The detailed FAC analysis further implied that the presence of terminal GlcNAc at the N-acetylglucosaminyltransferase I position is a key determinant for the binding of these lectins to agalactosylated N-glycans. By .