The trafficking of prosaposin (SGP-1) and GM2AP to the lysosomes of TM4 Sertoli cells is mediated by sortilin and monomeric adaptor proteins.
Prosaposin (SGP-1) and GM2 activator protein (GM2AP) are soluble sphingolipid activator proteins (SAPs) that are targeted to the lysosomal compartment of Sertoli cells to aid hydrolases in the breakdown of glycosphingolipids. To reach the lysosome, most soluble proteins must interact with the mannose 6-phosphate receptor (MPR). To be sorted from the Golgi, the MPR must bind to the Golgi associated, gamma-adaptin homologous, ARF binding proteins (GGAs), a group of monomeric adaptor proteins responsible for the recruitment of clathrin. It is well established, however, that the lysosomes of I-cell disease (ICD) patients have near normal levels of several lysosomal proteins, including prosaposin and GM2AP. ICD results from a mutation in the phosphotransferase that adds mannose 6-phosphate to hydrolases. Thus, prosaposin and GM2AP can traffic to lysosomes in a MPR independent manner. Previous work has demonstrated that an interaction with sphingomyelin in the Golgi membrane is necessary for the targeting of prosaposin by an unknown receptor. Using a TM4 Sertoli cell line, we tested the hypothesis that prosaposin and GM2AP are targeted to the lysosomal compartment via the sortilin receptor, which has been recently shown to have a GGA binding motif. Interestingly, dominant-negative GGAs, unable to bind clathrin to shuttle from the Golgi, prevented the trafficking of prosaposin and GM2AP to lysosomes. A dominant negative construct of sortilin lacking the GGA binding domain retained prosaposin and GM2AP in the Golgi. In conclusion, our results showed that the trafficking of prosaposin and GM2AP to the lysosome is dependent on sortilin.
Hassan
A Jacob
AJ
Department of Anatomy and Cell Biology, McGill University, Montreal, Quebec, Canada.
Zeng
Jibin
J
Ni
Xiaoyan
X
Morales
Carlos R
CR
D033942:Adaptor Proteins, Vesicular Transport; D000818:Animals; D049289:G(M2) Activator Protein / Q000235:genetics*; D017930:Genes, Reporter; D006056:Golgi Apparatus / Q000502:physiology; D049452:Green Fluorescent Proteins / Q000235:genetics; D008247:Lysosomes / Q000502:physiology*; D008297:Male; D008562:Membrane Glycoproteins / Q000235:genetics; D051379:Mice; D009419:Nerve Tissue Proteins / Q000235:genetics; D049231:Saposins / Q000502:physiology*; D012708:Sertoli Cells / Q000502:physiology*; D000099183:Sortilin
D033942:Adaptor Proteins, Vesicular Transport; D049289:G(M2) Activator Protein; D049452:Green Fluorescent Proteins; D008562:Membrane Glycoproteins; D009419:Nerve Tissue Proteins; D049231:Saposins; D000099183:Sortilin; C483344:Psap protein, mouse