Vacuolar Protein Sorting 36 (VPS36)

Eap30 (SNF8) and Eap20 (VPS25) could be coimmunoprecipitated in the absence of Eap45, and that Eap20 and Eap45 could be coimmunoprecipitated in the absence of Eap30. However, little Eap30 was coimmunoprecipitated with Eap45 in the absence of Eap20. The Eap45 GLUE domain shares primary and secondary structures with those of the phosphoinositide-binding GRAM and pleckstrin (PLEK) homology (PH) domains.GLUE domain of Eap45 bound ubiquitin with similar affinity and specificity as other ubiquitin-binding domains. The GLUE domain of Eap45 also bound specifically to phosphatidylinositol 3,4,5-trisphosphate (PtdIns(3,4,5)P3) and more weakly to PtdIns(3,4)P2 and PtdIns(3,5)P2. Eap45 colocalized with ubiquitinated proteins on late endosomes.