title: Phosphoinositides differentially regulate protrudin localization through the FYVE domain.
abstract: Protrudin is a FYVE (Fab 1, YOTB, Vac 1, and EEA1) domain-containing protein involved in transport of neuronal cargoes and implicated in the onset of hereditary spastic paraplegia. Our image-based screening of the lipid binding domain library revealed novel plasma membrane localization of the FYVE domain of protrudin unlike canonical FYVE domains that are localized to early endosomes. The membrane binding study by surface plasmon resonance analysis showed that this FYVE domain preferentially binds phosphatidylinositol 4,5-bisphosphate (PtdIns(4,5)P(2)), phosphatidylinositol 3,4-bisphosphate (PtdIns(3,4)P(2)), and phosphatidylinositol 3,4,5-trisphosphate (PtdIns(3,4,5)P(3)) unlike canonical FYVE domains that specifically bind phosphatidylinositol 3-phosphate (PtdIns(3)P). Furthermore, we found that these phosphoinositides (PtdInsP) differentially regulate shuttling of protrudin between endosomes and plasma membrane via its FYVE domain. Protrudin mutants with reduced PtdInsP-binding affinity failed to promote neurite outgrowth in primary cultured hippocampal neurons. These results suggest that novel PtdInsP selectivity of the protrudin-FYVE domain is critical for its cellular localization and its role in neurite outgrowth.
authors.affiliation: Department of Biological Sciences, Korea Advanced Institute of Science and Technology, Daejeon 305-701, Republic of Korea.
authors.forename: Jung-Eun
authors.initials: JE
authors.lastname: Gil
authors.forename: Eui
authors.initials: E
authors.lastname: Kim
authors.forename: Il-Shin
authors.initials: IS
authors.lastname: Kim
authors.forename: Bonsu
authors.initials: B
authors.lastname: Ku
authors.forename: Wei Sun
authors.initials: WS
authors.lastname: Park
authors.forename: Byung-Ha
authors.initials: BH
authors.lastname: Oh
authors.forename: Sung Ho
authors.initials: SH
authors.lastname: Ryu
authors.forename: Wonhwa
authors.initials: W
authors.lastname: Cho
authors.forename: Won Do
authors.initials: WD
authors.lastname: Heo
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D002352:Carrier Proteins / Q000096:biosynthesis* / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D002462:Cell Membrane / Q000378:metabolism; D011992:Endosomes / Q000378:metabolism; D007700:Kinetics; D008055:Lipids / Q000737:chemistry; D051379:Mice; D008954:Models, Biological; D008958:Models, Molecular; D008969:Molecular Sequence Data; D009154:Mutation; D041681:NIH 3T3 Cells; D016501:Neurites / Q000378:metabolism; D018129:Phosphatidylinositol Phosphates / Q000737:chemistry*; D010716:Phosphatidylinositols / Q000378:metabolism*; D011487:Protein Conformation; D017434:Protein Structure, Tertiary; D017386:Sequence Homology, Amino Acid; D020349:Surface Plasmon Resonance / Q000379:methods; D033921:Vesicular Transport Proteins
chemical_list: D002352:Carrier Proteins; D008055:Lipids; D018129:Phosphatidylinositol Phosphates; D010716:Phosphatidylinositols; D033921:Vesicular Transport Proteins; C055525:phosphatidylinositol 3-phosphate; C514783:protrudin protein, mouse