Synthesis and secretion of lipoprotein lipase in heparan sulfate-deficient Chinese hamster ovary cells.
Synthesis and secretion of lipoprotein lipase was studied in two mutants of Chinese hamster ovary (CHO) cells which, due to a lack of xylosyl transferase (pgsA-745) or galactosyl transferase (pgsB-761), respectively, were deficient in heparan sulfate and chondroitin sulfate. One of the mutants (pgsB-761) was two- to threefold more active in synthesis and secretion of catalytically active lipoprotein lipase than the other mutant, which was about as active as the wild-type (K1) cells. A similar relation was found when lipoprotein lipase was metabolically labelled with 35S-methionine and then immunoprecipitated. Heparin stimulated secretion from all three cell types to a similar extent (about twofold). Heparin-releasable binding of 125I-labelled lipoprotein lipase was lower to either of the mutant cells than to the wild-type cells. Binding to the wild-type cells was reduced by heparitinase, while the low binding to the mutants was not affected. By immunogold labelling of cryosections, lipoprotein lipase was detected on the plasma membranes and on the inside of secretory vesicles of both wild-type and mutant cells, suggesting that some carrier could be involved. Inhibition of vesicular transport by monensin caused accumulation of lipoprotein lipase in the cells. In wild-type cells the lipase was mainly on the inside of vesicular structures, while in the mutants the main part was associated with membranous bodies that formed within the vesicles during a chase period. These results suggest that if lipoprotein lipase needs a carrier during intracellular assembly and transport, this function can be fulfilled by some structure other than heparan sulfate.
Olivecrona
G
G
Department of Medical Biochemistry and Biophysics, Umeå University, Sweden.
Vilaró
S
S
Esko
J D
JD
Olivecrona
T
T
D000818:Animals; D016466:CHO Cells; D002809:Chondroitin Sulfates / Q000172:deficiency; D006224:Cricetinae; D003513:Cycloheximide / Q000494:pharmacology; D006493:Heparin / Q000494:pharmacology; D006497:Heparan Sulfate / Q000172:deficiency*; D008071:Lipoprotein Lipase / Q000096:biosynthesis* / Q000378:metabolism; D016253:Microscopy, Immunoelectron; D008985:Monensin / Q000494:pharmacology; D011133:Polysaccharide-Lyases / Q000494:pharmacology; D011500:Protein Synthesis Inhibitors / Q000494:pharmacology
D002809:Chondroitin Sulfates; D003513:Cycloheximide; D006493:Heparin; D006497:Heparan Sulfate; D008071:Lipoprotein Lipase; D008985:Monensin; D011133:Polysaccharide-Lyases; D011500:Protein Synthesis Inhibitors; C021349:heparitinsulfate lyase