title: Cytotoxic amphiphiles and phosphoinositides bind to two discrete sites on the Akt1 PH domain.
abstract: The mechanism of binding of two promising anticancer agents (the cytotoxic alkylphospholipids perifosine and miltefosine) to the Akt PH domain is investigated by high-resolution field-cycling (31)P nuclear magnetic resonance (NMR) spectroscopy using a spin-labeled recombinant PH domain. These results strongly indicate that there are two discrete amphiphile binding sites on the domain: (i) the cationic site that binds phosphoinositides and some alkylphospholipids and (ii) a second site that is occupied by only the alkylphospholipids. The identification of this second site for amphiphiles on the Akt1 PH domain provides a new target for drug development as well as insights into the regulation of the activity of the intact Akt1 protein. The field-cycling NMR methodology could be used to define discrete phospholipid or amphiphile binding sites on a wide variety of peripheral membrane proteins.
authors.affiliation: Department of Chemistry, Boston College , Chestnut Hill, Massachusetts 02467, United States.
authors.forename: Cheryl S
authors.initials: CS
authors.lastname: Gradziel
authors.forename: Yanling
authors.initials: Y
authors.lastname: Wang
authors.forename: Boguslaw
authors.initials: B
authors.lastname: Stec
authors.forename: Alfred G
authors.initials: AG
authors.lastname: Redfield
authors.forename: Mary F
authors.initials: MF
authors.lastname: Roberts
mesh_terms: D000970:Antineoplastic Agents / Q000378:metabolism; D001665:Binding Sites; D006801:Humans; D008823:Micelles; D008958:Models, Molecular; D019906:Nuclear Magnetic Resonance, Biomolecular; D010716:Phosphatidylinositols / Q000378:metabolism*; D010767:Phosphorylcholine / Q000031:analogs & derivatives* / Q000378:metabolism; D017434:Protein Structure, Tertiary; D051057:Proto-Oncogene Proteins c-akt / Q000737:chemistry / Q000378:metabolism*; D013113:Spin Labels
chemical_list: D000970:Antineoplastic Agents; D008823:Micelles; D010716:Phosphatidylinositols; D013113:Spin Labels; D010767:Phosphorylcholine; C105905:perifosine; C039128:miltefosine; D051057:Proto-Oncogene Proteins c-akt