RESUMEN
Immunoglobulin M (IgM) is the first antibody to emerge during embryonic development and the humoral immune response1. IgM can exist in several distinct forms, including monomeric, membrane-bound IgM within the B cell receptor (BCR) complex, pentameric and hexameric IgM in serum and secretory IgM on the mucosal surface. FcµR, the only IgM-specific receptor in mammals, recognizes different forms of IgM to regulate diverse immune responses2-5. However, the underlying molecular mechanisms remain unknown. Here we delineate the structural basis of the FcµR-IgM interaction by crystallography and cryo-electron microscopy. We show that two FcµR molecules interact with a Fcµ-Cµ4 dimer, suggesting that FcµR can bind to membrane-bound IgM with a 2:1 stoichiometry. Further analyses reveal that FcµR-binding sites are accessible in the context of IgM BCR. By contrast, pentameric IgM can recruit four FcµR molecules to bind on the same side and thereby facilitate the formation of an FcµR oligomer. One of these FcµR molecules occupies the binding site of the secretory component. Nevertheless, four FcµR molecules bind to the other side of secretory component-containing secretory IgM, consistent with the function of FcµR in the retrotransport of secretory IgM. These results reveal intricate mechanisms of IgM perception by FcµR.
Asunto(s)
Proteínas Reguladoras de la Apoptosis , Inmunoglobulina M , Proteínas de la Membrana , Animales , Linfocitos B/citología , Linfocitos B/metabolismo , Sitios de Unión , Membrana Celular/metabolismo , Microscopía por Crioelectrón , Cristalografía por Rayos X , Inmunoglobulina M/química , Inmunoglobulina M/metabolismo , Inmunoglobulina M/ultraestructura , Mamíferos , Unión Proteica , Multimerización de Proteína , Receptores de Antígenos de Linfocitos B/química , Receptores de Antígenos de Linfocitos B/metabolismo , Receptores de Antígenos de Linfocitos B/ultraestructura , Componente Secretorio/química , Componente Secretorio/metabolismo , Componente Secretorio/ultraestructura , Proteínas de la Membrana/química , Proteínas de la Membrana/metabolismo , Proteínas de la Membrana/ultraestructura , Proteínas Reguladoras de la Apoptosis/química , Proteínas Reguladoras de la Apoptosis/metabolismo , Proteínas Reguladoras de la Apoptosis/ultraestructuraRESUMEN
Immunogold labeling was applied at the ultrastructural level to the identification of collagen types I, II, III, V, and VI in the human trabecular meshwork obtained from 11 surgically enucleated globes. Both London resin white embedding and cryoultramicrotomy were used, and for types V and VI, the latter provided more sensitive localization. Type II collagen was not identified. Types I and III were localized to the striated collagen fibrils of the trabecular core, the basement membrane of the trabecular beams, and loose aggregates in the juxtacanalicular tissue. Types V and VI formed a fine network around the striated fibrils in the trabecular cores and linkage strands to the basement membranes. The outer wall of Schlemm's canal contained collagens I, III, V, and VI. Long-spacing collagen and elastin-like material did not label with any of the antibodies used in this study.