Glycosylation, transport, and complex formation of palmitoyl protein thioesterase 1 (PPT1) – distinct characteristics in neurons-10

<p><b>Copyright information:</b></p><p>Taken from "Glycosylation, transport, and complex formation of palmitoyl protein thioesterase 1 (PPT1) – distinct characteristics in neurons"</p><p>http://www.biomedcentral.com/1471-2121/8/22</p><p>BMC Cell Biology 2007;8():22-22.</p><p>Published online 12 Jun 2007</p><p>PMCID:PMC1906764.</p><p></p>+ S214A. The cells were double-stained using the polyclonal antibody for PPT1 (red) and the monoclonal antibody for LAMP-1, lysosomal membrane protein (green). Colocalization is shown in yellow

Glycosylation, transport, and complex formation of palmitoyl protein thioesterase 1 (PPT1) – distinct characteristics in neurons-8

<p><b>Copyright information:</b></p><p>Taken from "Glycosylation, transport, and complex formation of palmitoyl protein thioesterase 1 (PPT1) – distinct characteristics in neurons"</p><p>http://www.biomedcentral.com/1471-2121/8/22</p><p>BMC Cell Biology 2007;8():22-22.</p><p>Published online 12 Jun 2007</p><p>PMCID:PMC1906764.</p><p></p>h GFP-vector (c) or with the corresponding GFP-PPT1 construct having the same mutation in the PPT molecule (m). The cells were lysed 48 h post-transfection. To verify the transfections, 10 μg of total protein from the lysates was analyzed by SDS-PAGE and immunoblotting using both anti-GFP antibody (not shown) and anti-PPT1 antibody (Transfection). The lysates were then subjected to immunoprecipitation using anti-GFP-agarose beads. Immunoprecipitates were analyzed by SDS-PAGE and immunoblotting using the anti-PPT1 antibody (IP). The graph represents the dimerization degree of mutant proteins compared to wild type (calculated as a ratio between the immunoprecipitated GFP-PPT/PPT and the transfected GFP-PPT/PPT). An example of a single experiment's quantification is shown experiments

Glycosylation, transport, and complex formation of palmitoyl protein thioesterase 1 (PPT1) – distinct characteristics in neurons-6

<p><b>Copyright information:</b></p><p>Taken from "Glycosylation, transport, and complex formation of palmitoyl protein thioesterase 1 (PPT1) – distinct characteristics in neurons"</p><p>http://www.biomedcentral.com/1471-2121/8/22</p><p>BMC Cell Biology 2007;8():22-22.</p><p>Published online 12 Jun 2007</p><p>PMCID:PMC1906764.</p><p></p>ned for PPT1 using the GST-PPT1 antibody (green), for lysosomes using the Lamp-1 antibody (red), for synaptic vesicles using the SV2 antibody (red) and for ER using the PDI antibody (red). Colocalization is shown in yellow. Scale bar = 20 μm

Glycosylation, transport, and complex formation of palmitoyl protein thioesterase 1 (PPT1) – distinct characteristics in neurons-3

<p><b>Copyright information:</b></p><p>Taken from "Glycosylation, transport, and complex formation of palmitoyl protein thioesterase 1 (PPT1) – distinct characteristics in neurons"</p><p>http://www.biomedcentral.com/1471-2121/8/22</p><p>BMC Cell Biology 2007;8():22-22.</p><p>Published online 12 Jun 2007</p><p>PMCID:PMC1906764.</p><p></p>ion was subjected to size exclusion chromatography and the PPT1 and AGA enzyme activities were measured from the fractions collected. The X-axis indicates time and the molecular weights are derived from the standard (Bio-Rad)

Glycosylation, transport, and complex formation of palmitoyl protein thioesterase 1 (PPT1) – distinct characteristics in neurons-0

<p><b>Copyright information:</b></p><p>Taken from "Glycosylation, transport, and complex formation of palmitoyl protein thioesterase 1 (PPT1) – distinct characteristics in neurons"</p><p>http://www.biomedcentral.com/1471-2121/8/22</p><p>BMC Cell Biology 2007;8():22-22.</p><p>Published online 12 Jun 2007</p><p>PMCID:PMC1906764.</p><p></p> each lysate (5 μg total protein) was analyzed by a one-hour reaction in + 37°C. Transfection efficacies were controlled by immunofluorescence analysis. The data shown represents an average of three independent experiments and the standard deviations between these experiments are marked as error bars. COS-1 cells were transfected and metabolically labelled with 35S-Cysteine for 1 hour, chased for 2 hours, immunoprecipitated, separated in SDS-PAGE and autoradiographed. Both intracellular and secreted forms of PPT are shown

Glycosylation, transport, and complex formation of palmitoyl protein thioesterase 1 (PPT1) – distinct characteristics in neurons-1

<p><b>Copyright information:</b></p><p>Taken from "Glycosylation, transport, and complex formation of palmitoyl protein thioesterase 1 (PPT1) – distinct characteristics in neurons"</p><p>http://www.biomedcentral.com/1471-2121/8/22</p><p>BMC Cell Biology 2007;8():22-22.</p><p>Published online 12 Jun 2007</p><p>PMCID:PMC1906764.</p><p></p> + S214A. The cells were double-stained using the polyclonal antibody for PPT1 (red) and the monoclonal antibody for LAMP-1, lysosomal membrane protein (green). Colocalization is shown in yellow

Glycosylation, transport, and complex formation of palmitoyl protein thioesterase 1 (PPT1) – distinct characteristics in neurons-7

<p><b>Copyright information:</b></p><p>Taken from "Glycosylation, transport, and complex formation of palmitoyl protein thioesterase 1 (PPT1) – distinct characteristics in neurons"</p><p>http://www.biomedcentral.com/1471-2121/8/22</p><p>BMC Cell Biology 2007;8():22-22.</p><p>Published online 12 Jun 2007</p><p>PMCID:PMC1906764.</p><p></p>d densitometric scanning. The numbers 3, 2 and 1 indicate the three differently glycosylated forms of PPT1. The graph represents relative amounts of tri-, di-, and monoglycosylated forms of PPT1 in transfected COS-1 cells. The data shown represents an average of three independent experiments and the error bars show the standard deviations between these experiments

Adenovirus-mediated PPT1 was expressed in mouse fibroblasts and neurons

<p><b>Copyright information:</b></p><p>Taken from "Glycosylation, transport, and complex formation of palmitoyl protein thioesterase 1 (PPT1) – distinct characteristics in neurons"</p><p>http://www.biomedcentral.com/1471-2121/8/22</p><p>BMC Cell Biology 2007;8():22-22.</p><p>Published online 12 Jun 2007</p><p>PMCID:PMC1906764.</p><p></p> Cell lysates (5 μg of total protein) were treated with PNGaseF to remove the N-glycans from the polypeptides. Proteins were separated with SDS-PAGE and analyzed by immunoblotting. Antibody internalization assay was performed in adenovirus infected mouse fibroblasts and neurons with both AGA and PPT1 (green). Double staining with the lysosomal marker LAMP-1 (red) is shown for internalized PPT1 in fibroblasts . Co-localization is shown in yellow

Western blot analysis of cytoplasmic and membrane bound fractions of cytoskeletal, growth-cone and synapse assembly proteins

Antibodies: β-tubulin, synapsin 1, synapsin 1&2, Rab3 and Gap43.<p><b>Copyright information:</b></p><p>Taken from "Brain gene expression profiles of and deficient mice unravels common molecular pathways underlying neuronal degeneration in NCL diseases"</p><p>http://www.biomedcentral.com/1471-2164/9/146</p><p>BMC Genomics 2008;9():146-146.</p><p>Published online 28 Mar 2008</p><p>PMCID:PMC2323392.</p><p></p

Several differentially expressed genes show significant clustering in around on mouse chromosome 4, and might therefore be coregulated

The probesets for Ppt1 were excluded from the dataset and thus Ppt1 is shown in gray. The genes participating in the regulated loci or being independently regulated are shown, together with their location in base pairs in mouse chromosome 4. Genes that exhibit expression differences in both mouse models are indicated with *. Expression changes only in the mouse are indicated with **.<p><b>Copyright information:</b></p><p>Taken from "Brain gene expression profiles of and deficient mice unravels common molecular pathways underlying neuronal degeneration in NCL diseases"</p><p>http://www.biomedcentral.com/1471-2164/9/146</p><p>BMC Genomics 2008;9():146-146.</p><p>Published online 28 Mar 2008</p><p>PMCID:PMC2323392.</p><p></p