Localization of lysosomal antigens in activated T-lymphocytes.
The lysosomal compartment has been examined in activated T-lymphocytes by immunogold electron microscopy and subcellular fractionation. Immunoprecipitation and sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE) of radiolabelled extracts of the T-cells showed that they contained three antigens which are fundamental to normal lysosomal function: a representative lysosomal enzyme beta-glucuronidase, a lysosomal associated membrane protein (LAMP-1), and the cation-independent mannose 6-phosphate lysosomal enzyme targeting receptor (MPR). Immunogold labelling showed that beta-glucuronidase was present in the rough endoplasmic reticulum, the Golgi complex and Golgi-associated vesicles. The enzyme was also found to accumulate in distinct, non-Golgi organelles in which LAMP-1 was co-localized, probably lysosomes. LAMP-1 was also found in tubular elements of the Golgi and in a complex of vesicles clustered near the nucleus where MPR was also present at high density. Fractionation of homogenates from lymphocytes on Percoll gradients revealed that beta-glucuronidase was distributed throughout the low density region containing rough endoplasmic reticulum, Golgi and plasma membrane components, and the high density region which contained only lysosomal activity. Multiple immunogold electron microscopy of the latter fraction showed the presence of homogenous vesicles which had large amounts of beta-glucuronidase within the lumen, LAMP-1 at the periphery and no MPR. These vesicles were probably mature lysosomes, arising from pre-lysosomal organelles enriched for LAMP-1 and MPR.
Bou-Gharios
G
G
Cell Enzymology Unit, Kennedy Institute of Rheumatology, London, UK.
Moss
J
J
Olsen
I
I
D000818:Animals; D000941:Antigens / Q000032:analysis*; D004591:Electrophoresis, Polyacrylamide Gel; D005966:Glucuronidase / Q000032:analysis; D008247:Lysosomes / Q000276:immunology*; D051379:Mice; D008808:Mice, Inbred CBA; D016253:Microscopy, Immunoelectron; D011233:Precipitin Tests; D013154:Spleen / Q000276:immunology; D013601:T-Lymphocytes / Q000276:immunology*; D014018:Tissue Distribution
D000941:Antigens; D005966:Glucuronidase