title: Membrane topology of helix 0 of the Epsin N-terminal homology domain.
abstract: Specific interaction of the epsin N-terminal homology (ENTH) domain with the plasma membrane appears to bridge other related proteins to the specific regions of the membrane that are invaginated to form endocytic vesicles. An additional a-helix, referred to as helix 0 (H0), is formed in the presence of the soluble ligand inositol-1,4,5-trisphosphate [Ins(1,4,5)P3] at the N terminus of the ENTH domain (amino acid residues 3-15). The ENTH domain alone and full-length epsin cause tubulation of liposomes made of brain lipids. Thus, it is believed that H0 is membrane-inserted when it is coordinated with the phospholipid phosphatidylinositol-4,5-bisphosphate [PtdIns(4,5)P2], resulting in membrane deformation as well as recruitment of accessory factors to the membrane. However, formation of H0 in a real biological membrane has not been demonstrated. In the present study, the membrane structure of H0 was determined by measurement of electron paramagnetic resonance (EPR) nitroxide accessibility. H0 was located at the phosphate head-group region of the membrane. Moreover, EPR line-shape analysis indicated that no pre-formed H0-like structure were present on normal acidic membranes. PtdIns(4,5)P2 was necessary and sufficient for interaction of the H0 region with the membrane. H0 was stable only in the membrane. In conclusion, the H0 region of the ENTH domain has an intrinsic ability to form H0 in a PtdIns(4,5)P2-containing membrane, perhaps functioning as a sensor of membrane patches enriched with PtdIns(4,5)P2 that will initiate curvature to form endocytic vesicles.
authors.affiliation: Department of Genetic Engineering, Faculty of Life Science and Technology, Sungkyunkwan University, Suwon 440-746, Korea. dhkweon @skku.edu
authors.forename: Dae-Hyuk
authors.initials: DH
authors.lastname: Kweon
authors.forename: Yeon-Kyun
authors.initials: YK
authors.lastname: Shin
authors.forename: Jae Yoon
authors.initials: JY
authors.lastname: Shin
authors.forename: Jong-Hwa
authors.initials: JH
authors.lastname: Lee
authors.forename: Jung-Bok
authors.initials: JB
authors.lastname: Lee
authors.forename: Jin-Ho
authors.initials: JH
authors.lastname: Seo
authors.forename: Yong Sung
authors.initials: YS
authors.lastname: Kim
mesh_terms: D033942:Adaptor Proteins, Vesicular Transport / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D000818:Animals; D001665:Binding Sites; D002462:Cell Membrane / Q000737:chemistry* / Q000378:metabolism; D004578:Electron Spin Resonance Spectroscopy; D008081:Liposomes; D016297:Mutagenesis, Site-Directed; D019269:Phosphatidylinositol 4,5-Diphosphate / Q000737:chemistry* / Q000378:metabolism; D017434:Protein Structure, Tertiary; D051381:Rats
chemical_list: D033942:Adaptor Proteins, Vesicular Transport; D008081:Liposomes; D019269:Phosphatidylinositol 4,5-Diphosphate; C114154:epsin