Serglycin and secretion in human monocytes.
The human monocytic cell line U-937 has been widely used as a model system for human monocytes. The subclone U-937-B has been adapted to serum-free conditions. This particular U-937 clone and its parent clone U-937-1 were used to investigate the role of the proteoglycan serglycin in human monocytes. For this purpose cells were treated with hexyl-beta-D-thioxyloside to abrogate proteoglycan expression. U-937-B cells expressed and secreted exclusively chondroitin sulphate proteoglycans, and after treatment with this xyloside they only expressed and released free chondroitin sulphate chains. Western blotting showed that serglycin core protein was present in conditioned medium of control cells, but absent in medium from xyloside-treated cells. Also, serglycin core protein could be detected in the cell fractions of control cells, but not in the cell fractions from xyloside-treated cells. Furthermore, less proteoglycan-associated proteins could be detected in medium from cells incubated with xyloside, suggesting that the absence of secreted sergycin affects the secretion of such proteins. Cells incubated in the presence of xyloside were analyzed by transmission electron microscopy and shown to contain numerous large empty vesicles. The lack of serglycin, the dominant proteoglycan in U-937 monocyte-like cells, consequently, leads to effects on vesicle formation and secretion of some low molecular weight proteins, suggesting that this particular proteoglycan is of importance for secretory processes in human monocytes.
Kolset
Svein Olav
SO
Department of Nutrition, Institute of Basic Medical Sciences, University of Oslo, Box 1046, Blindern, 0316, Oslo, Norway. s.o.kolset@medisin.uio.no
Zernichow
Lillian
L
D015153:Blotting, Western; D049109:Cell Proliferation; D002846:Chromatography, Affinity; D002852:Chromatography, Ion Exchange; D017077:Culture Media, Conditioned; D004591:Electrophoresis, Polyacrylamide Gel; D006801:Humans; D009000:Monocytes / Q000166:cytology / Q000378:metabolism* / Q000648:ultrastructure; D011509:Proteoglycans / Q000378:metabolism*; D013347:Subcellular Fractions / Q000378:metabolism; D020298:U937 Cells; D033921:Vesicular Transport Proteins / Q000378:metabolism*
D017077:Culture Media, Conditioned; D011509:Proteoglycans; D033921:Vesicular Transport Proteins; C075430:serglycin