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Rat Brain Contains High Levels of Mannose-6-phosphorylated Glycoproteins Including Lysosomal Enzymes and Palmitoyl-Protein Thioesterase, an Enzyme Implicated in Infantile Neuronal Lipofuscinosis

Mannose 6-phosphate (Man-6-P) is a posttranslational carbohydrate modification typical of newly synthesized acid hydrolases that signals targeting from the Golgi apparatus to the lysosome via Man-6-P receptors (MPRs). Using iodinated cation independent MPR as a probe in a Western blot assay, we surv...

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Bibliographic Details
Published in:The Journal of biological chemistry 1996-08, Vol.271 (32), p.19191-19198
Main Authors: Sleat, D E, Sohar, I, Lackland, H, Majercak, J, Lobel, P
Format: Article
Language:English
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Summary:Mannose 6-phosphate (Man-6-P) is a posttranslational carbohydrate modification typical of newly synthesized acid hydrolases that signals targeting from the Golgi apparatus to the lysosome via Man-6-P receptors (MPRs). Using iodinated cation independent MPR as a probe in a Western blot assay, we surveyed levels of Man-6-P glycoproteins in a number of different rat tissues. Considerable variation was observed with respect to total amounts and types of Man-6-P glycoproteins in the different tissues. Brain contained 2-8-fold more Man-6-P glycoproteins than other tissues, with relative abundance being brain ≫ testis ≈ heart > lung ≈ kidney ≈ ovary ≈ spleen > skeletal muscle ≈ liver ≈ serum. Analysis of 16 different lysosomal enzyme activities revealed that brain contains lower activities than other tissues which suggested that decreased removal of Man-6-P results in increased levels of Man-6-P glycoproteins. This was directly demonstrated by comparing activities of phosphorylated lysosomal enzymes, purified by immobilized MPR affinity chromatography, with total activities. The phosphorylated forms accounted for a considerable proportion of the MPR-targeted activities measured in brain (on average, 36.2%) but very little in lung, kidney, and liver (on average, 5.5, 2.3, and 0.7%, respectively). Man-6-P glycoproteins were also isolated from rat brain by MPR affinity chromatography on a preparative scale. Of the 18 bands resolvable by SDS-polyacrylamide gel electrophoresis, seven bands were NH 2 -terminally sequenced and identified as the known lysosomal enzymes cathepsin L, cathepsin A, cathepsin D, α-galactosidase A, arylsulfatase A, and α-iduronidase. One of the major Man-6-P glycoproteins was identified as palmitoyl protein thioesterase, which was not previously thought to be lysosomal. This finding raises important questions about the cellular location and function of palmitoyl protein thioesterase, mutations in which result in the neurodegenerative disorder, infantile neuronal ceroid lipofuscinosis.
ISSN:0021-9258
1083-351X
DOI:10.1074/jbc.271.32.19191