title: A novel calcium-independent peripheral membrane-bound form of annexin B12.
abstract: Annexins are soluble proteins that can interact with membranes in a Ca2+-dependent manner. Recent studies have shown that they can also undergo Ca2+-independent membrane interactions that are modulated by pH and phospholipid composition. Here, we investigated the structural changes that occurred during Ca2+-independent interaction of annexin B12 with phospholipid vesicles as a function of pH. Electron paramagnetic resonance analysis of a helical hairpin encompassing the D and E helices in the second repeat of the protein showed that this region refolded and formed a continuous amphipathic alpha helix following Ca2+-independent binding to membranes at mildly acidic pH. At pH 4.0, this helix assumed a transmembrane topography, but at pH approximately 5.0-5.5, it was peripheral and approximately parallel to the membrane. The peripheral form was reversibly converted into the transmembrane form by lowering the pH and vice versa. Furthermore, analysis of vesicles incubated with annexin B12 using freeze-fracture electron microscopy methods showed classical intramembrane particles at pH 4.0 but none at pH 5.3. Together, these data raise the possibility that the peripheral-bound form of annexin B12 could act as a kinetic intermediate in the formation of the transmembrane form of the protein.
authors.affiliation: Department of Biochemistry and Molecular Biology, Zilkha Neurogenetic Institute, University of Southern California, Keck School of Medicine, Los Angeles, California 90033, USA.
authors.forename: Balachandra G
authors.initials: BG
authors.lastname: Hegde
authors.forename: J Mario
authors.initials: JM
authors.lastname: Isas
authors.forename: Guido
authors.initials: G
authors.lastname: Zampighi
authors.forename: Harry T
authors.initials: HT
authors.lastname: Haigler
authors.forename: Ralf
authors.initials: R
authors.lastname: Langen
mesh_terms: D017302:Annexins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D002118:Calcium / Q000378:metabolism; D002462:Cell Membrane / Q000378:metabolism*; D005614:Freeze Fracturing; D006863:Hydrogen-Ion Concentration; D008958:Models, Molecular; D011485:Protein Binding
chemical_list: D017302:Annexins; D002118:Calcium