|
|
|
|
|
|
|
||
The Journal of Cell Biology, Vol 93, 875-882, Copyright © 1982 by The Rockefeller University Press
ARTICLES |
MC Kielian and ZA Cohn
Dextran sulfate (DS) was previously shown to inhibit phagosome-lysosome (P-L) fusion whereas dextran (D) of equivalent size was ineffective. The uptake and interiorization of DS were examined with a tritiated product over the course of 4 d in culture. The exposure of macrophages to 20 micrograms/ml of 3H-DS led to linear uptake for 4 d, at which time fusion was inhibited. Macrophage interiorization of 3H-DS was greatly increased by forming insoluble complexes with either serum lipoproteins or purified human low density lipoproteins (LDL). Under these conditions fusion was inhibited within 4 h. The uptake of large quantities of acetylated LDL in the absence of DS was not associated with the inhibition of fusion. Lipoproteins therefore served as the DS carriers and were not themselves inhibitory. The intralysosomal pH of control and D-treated macrophages was 4.76 (+/-0.06) and 4.68 (+/- 0.02), respectively. Storage of DS was associated with a decreased pH to 4.36 (+/-0.14). Increasing the intralysosomal pH with either NH4Cl or chloroquine failed to modify inhibited P-L fusion. Hydrogen ion concentration was therefore not an important factor in DS inhibition. Secondary lysosomes were isolated from D- and DS-loaded cells and exhibited excellent latency. These lysosomes were exposed to the membrane probes, alpha- and Beta-parinaric acid, and compared in fluorescence polarization measurements. The results with the Beta isomer consistently indicated that the membranes of DS lysosomes were more rigid than the D samples. It is suggested that high intralysosomal concentrations of DS interact directly with either lipid and/or polypeptide moieties of the luminal face of the membrane, thereby decreasing its fluidity and fusibility.
CiteULike 
Complore 
Connotea 
Del.icio.us 
Digg 
Reddit 
Technorati What's this?
This article has been cited by other articles:
|
|
