title: AFM force spectroscopy reveals how subtle structural differences affect the interaction strength between Candida albicans and DC-SIGN.
abstract: The fungus Candida albicans is the most common cause of mycotic infections in immunocompromised hosts. Little is known about the initial interactions between Candida and immune cell receptors, such as the C-type lectin dendritic cell-specific intracellular cell adhesion molecule-3 (ICAM-3)-grabbing non-integrin (DC-SIGN), because a detailed characterization at the structural level is lacking. DC-SIGN recognizes specific Candida-associated molecular patterns, that is, mannan structures present in the cell wall of Candida. The molecular recognition mechanism is however poorly understood. We postulated that small differences in mannan-branching may result in considerable differences in the binding affinity. Here, we exploit atomic force microscope-based dynamic force spectroscopy with single Candida cells to gain better insight in the carbohydrate recognition capacity of DC-SIGN. We demonstrate that slight differences in the N-mannan structure of Candida, that is, the absence or presence of a phosphomannan side chain, results in differences in the recognition by DC-SIGN as follows: (i) it contributes to the compliance of the outer cell wall of Candida, and (ii) its presence results in a higher binding energy of 1.6 kB T. The single-bond affinity of tetrameric DC-SIGN for wild-type C. albicans is ~10.7 kB T and a dissociation constant kD of 23 μM, which is relatively strong compared with other carbohydrate-protein interactions described in the literature. In conclusion, this study shows that DC-SIGN specifically recognizes mannan patterns on C. albicans with high affinity. Knowledge on the binding pocket of DC-SIGN and its pathogenic ligands will lead to a better understanding of how fungal-associated carbohydrate structures are recognized by receptors of the immune system and can ultimately contribute to the development of new anti-fungal drugs.
authors.affiliation: Department of Tumor Immunology, Radboud Institute for Molecular Life Sciences, Radboud UMC, P.O. Box 9101, 6500HB, Nijmegen, The Netherlands.
authors.forename: Joost
authors.initials: J
authors.lastname: te Riet
authors.affiliation: Department of Tumor Immunology, Radboud Institute for Molecular Life Sciences, Radboud UMC, P.O. Box 9101, 6500HB, Nijmegen, The Netherlands.
authors.forename: Inge
authors.initials: I
authors.lastname: Reinieren-Beeren
authors.affiliation: Department of Tumor Immunology, Radboud Institute for Molecular Life Sciences, Radboud UMC, P.O. Box 9101, 6500HB, Nijmegen, The Netherlands.
authors.forename: Carl G
authors.initials: CG
authors.lastname: Figdor
authors.affiliation: Department of Tumor Immunology, Radboud Institute for Molecular Life Sciences, Radboud UMC, P.O. Box 9101, 6500HB, Nijmegen, The Netherlands.
authors.forename: Alessandra
authors.initials: A
authors.lastname: Cambi
mesh_terms: D001665:Binding Sites / Q000502:physiology; D002176:Candida albicans / Q000378:metabolism*; D002241:Carbohydrates; D002448:Cell Adhesion / Q000502:physiology; D015815:Cell Adhesion Molecules / Q000378:metabolism*; D002473:Cell Wall / Q000378:metabolism; D037181:Lectins, C-Type / Q000378:metabolism*; D008024:Ligands; D008351:Mannans / Q000378:metabolism; D018625:Microscopy, Atomic Force / Q000379:methods; D011485:Protein Binding / Q000502:physiology; D011956:Receptors, Cell Surface / Q000378:metabolism*; D013057:Spectrum Analysis / Q000379:methods; D000099215:DC-Specific ICAM-3 Grabbing Nonintegrin
chemical_list: D002241:Carbohydrates; D015815:Cell Adhesion Molecules; D037181:Lectins, C-Type; D008024:Ligands; D008351:Mannans; D011956:Receptors, Cell Surface; D000099215:DC-Specific ICAM-3 Grabbing Nonintegrin; C005437:phosphomannan
keywords: Candida albicans; DC-SIGN; atomic force microscopy; single molecule force spectroscopy; single-cell force spectroscopy