Title : Lysosomal targeting of the
CLN7 membrane
glycoprotein and transport via the plasma membrane require a dileucine
motif
Abstract :
- CLN7 is a polytopic lysosomal membrane protein deficient in variant late infantile neuronal ceroid lipofuscinosis, a neurodegenerative lysosomal storage disorder
- In this study fluorescence protease protection assays and mutational analyses revealed the N- and C-terminal tails of CLN7 in the cytosol and two N-glycosylation sites at N371 and N376
- Both partially and non-glycosylated CLN7 were correctly transported to lysosomes
- To identify lysosomal targeting motifs , we generated CD4-chimera fused to the N- and C-terminal domains of CLN7
- Lysosomal localization of the chimeric proteins requires a consensus acidic dileucine-based motif in the N-terminus and two tandem tyrosine-based signals in the C-terminus
- Mutation of these sorting motifs resulted in cell surface redistribution of CD4 chimeras
- However, the dileucine-based motif is of critical importance for lysosomal localization of the full-length CLN7 in different cell lines
- Cell surface biotinylation revealed that at equilibrium 22% of total CLN7 is localized at the plasma membrane
- Mutation of the dileucine motif or the co-expression of dominant-negative mutant dynamin K44A led to a further increase of CLN7 at the plasma membrane
- Our data demonstrate that CLN7 contains several cytoplasmic lysosomal targeting signals of which the N-terminal dileucine-based motif is required for the predominant lysosomal targeting along the indirect pathway and clathrin-mediated endocytosis of CLN7
Output (sent_index, trigger,
protein,
sugar,
site):
- 0. glycoprotein, , glycoprotein, -, -
- 2. N-glycosylation, , -, -, N371 and N376
- 2. N-glycosylation, , CLN7, -, N371 and N376
- 2. N-glycosylation, , CLN7, -, sites
- 3. non-glycosylated, , CLN7, -, -
Output(Part-Of) (sent_index,
protein,
site):
- 2. CLN7, N371 and N376
- 2. CLN7, sites
- 2. CLN7, tails
- 4. CLN7, domains
*Output_Site_Fusion* (sent_index,
protein,
sugar,
site):
- 2. CLN7, -, N371 and N376