Neutron reflectometry study of the conformation of HIV Nef bound to lipid membranes.
Nef is an HIV-1 accessory protein that directly contributes to AIDS progression. Nef is myristoylated on the N-terminus, associates with membranes, and may undergo a transition from a solution conformation to a membrane-associated conformation. It has been hypothesized that conformational rearrangement enables membrane-associated Nef to interact with cellular proteins. Despite its medical relevance, to our knowledge there is no direct information about the conformation of membrane-bound Nef. In this work, we used neutron reflection to reveal what we believe are the first details of the conformation of membrane-bound Nef. The conformation of Nef was probed upon binding to Langmuir monolayers through the interaction of an N-terminal His tag with a synthetic metal-chelating lipid, which models one of the possible limiting cases for myr-Nef. The data indicate that residues are inserted into the lipid headgroups during interaction, and that the core domain lies directly against the lipid headgroups, with a thickness of ∼40 A. Binding of Nef through the N-terminal His tag apparently facilitates insertion of residues, as no insertion occurred upon binding of Nef through weak electrostatic interactions in the absence of the specific interaction through the His tag.
Kent
Michael S
MS
Sandia National Laboratories, Albuquerque, New Mexico, USA. mskent@sandia.gov
Murton
Jaclyn K
JK
Sasaki
Darryl Y
DY
Satija
Sushil
S
Akgun
Bulent
B
Nanda
Hirsh
H
Curtis
Joseph E
JE
Majewski
Jaroslaw
J
Morgan
Christopher R
CR
Engen
John R
JR
D015060:1,2-Dipalmitoylphosphatidylcholine / Q000737:chemistry / Q000378:metabolism; D019342:Acetic Acid / Q000737:chemistry / Q000378:metabolism; D000327:Adsorption; D000595:Amino Acid Sequence; D002462:Cell Membrane / Q000737:chemistry / Q000378:metabolism*; D003300:Copper / Q000737:chemistry; D003903:Deuterium / Q000737:chemistry; D015497:HIV-1; D050356:Lipid Metabolism; D008958:Models, Molecular; D008969:Molecular Sequence Data; D033363:Neutron Diffraction / Q000379:methods*; D011485:Protein Binding; D011487:Protein Conformation; D021381:Protein Transport; D014961:X-Ray Diffraction; D054311:nef Gene Products, Human Immunodeficiency Virus / Q000737:chemistry* / Q000378:metabolism*
D054311:nef Gene Products, Human Immunodeficiency Virus; C514837:nef protein, Human immunodeficiency virus 1; D015060:1,2-Dipalmitoylphosphatidylcholine; D003300:Copper; D003903:Deuterium; D019342:Acetic Acid