Immunolocalization and molecular properties of a high molecular weight microtubule-bundling protein (syncolin) from chicken erythrocytes.
A protein of apparent molecular weight 280,000 (syncolin), which is immunoreactive with antibodies to hog brain microtubule-associated protein (MAP) 2, was purified from chicken erythrocytes. Immunofluorescence microscopy of bone marrow cells revealed the presence of syncolin in cells at all stages of erythrocyte differentiation. In early erythroblasts syncolin was diffusely distributed throughout the cytoplasm. At later stages it was found along microtubules of the marginal band, as confirmed by immunoelectron microscopy. The association of syncolin with the marginal band was dependent on the integrity of microtubules, as demonstrated by temperature-dependent de- and repolymerization or marginal band microtubules. Syncolin cosedimented in a saturable manner with microtubules assembled in vitro, and it was displaced from the polymer by salt. Brain as well as erythrocyte microtubules, reconstituted with taxol from MAP-free tubulin and purified syncolin, were aggregated into dense bundles containing up to 15 microtubules, as determined by electron microscopy. On the ultrastructural level, syncolin molecules were visualized as globular or ringlike structures, in contrast to the thin, threadlike appearance of filamentous MAPs, such as brain MAP 2. According to ultrastructural measurements and gel permeation chromatography, syncolin's molecular weight was approximately 1 x 10(6). It is suggested that syncolin's specific function is the cross-linking of microtubules in the marginal band and, by implication, the stabilization of this structure typical for nucleated (chicken) erythrocytes.
Feick
P
P
Institute of Biochemistry, University of Vienna, Austria.
Foisner
R
R
Wiche
G
G
D000818:Animals; D001798:Blood Proteins / Q000302:isolation & purification / Q000648:ultrastructure; D002645:Chickens; D004900:Erythroblasts / Q000737:chemistry / Q000648:ultrastructure; D004912:Erythrocytes / Q000737:chemistry* / Q000648:ultrastructure; D007150:Immunohistochemistry; D016253:Microscopy, Immunoelectron; D008869:Microtubule-Associated Proteins / Q000097:blood* / Q000302:isolation & purification / Q000648:ultrastructure; D008870:Microtubules / Q000378:metabolism
D001798:Blood Proteins; D008869:Microtubule-Associated Proteins