title: Mapping the interaction between the cytoplasmic domains of HIV-1 viral protein U and human CD4 with NMR spectroscopy.
abstract: Viral protein U (VpU) of HIV-1 plays an important role in downregulation of the main HIV-1 receptor CD4 from the surface of infected cells. Physical binding of VpU to newly synthesized CD4 in the endoplasmic reticulum is an early step in a pathway leading to proteasomal degradation of CD4. In this study, regions in the cytoplasmic domain of VpU involved in CD4 binding were identified by NMR spectroscopy. Amino acids in both helices found in the cytoplasmic region of VpU in membrane-mimicking detergent micelles experience chemical shift perturbations upon binding to CD4, whereas amino acids between the two helices and at the C-terminus of VpU show no or only small changes, respectively. The topology of the complex was further studied with paramagnetic relaxation enhancement. Paramagnetic spin labels were attached at three sequence positions of a CD4 peptide comprising the transmembrane and cytosolic domains of the receptor. VpU binds to a membrane-proximal region in the cytoplasmic domain of CD4.
authors.affiliation: Institute of Complex Systems (ICS-6), Forschungszentrum Jülich, Germany.
authors.forename: Sameer K
authors.initials: SK
authors.lastname: Singh
authors.affiliation: Institute of Complex Systems (ICS-6), Forschungszentrum Jülich, Germany.
authors.forename: Luis
authors.initials: L
authors.lastname: Möckel
authors.affiliation: Institute of Complex Systems (ICS-6), Forschungszentrum Jülich, Germany.
authors.forename: Pallavi
authors.initials: P
authors.lastname: Thiagarajan-Rosenkranz
authors.affiliation: Institute of Complex Systems (ICS-6), Forschungszentrum Jülich, Germany.
authors.forename: Marc
authors.initials: M
authors.lastname: Wittlich
authors.affiliation: Institute of Complex Systems (ICS-6), Forschungszentrum Jülich, Germany.|Institut für Physikalische Biologie, Heinrich-Heine-Universität Düsseldorf, Germany.
authors.forename: Dieter
authors.initials: D
authors.lastname: Willbold
authors.affiliation: Institute of Complex Systems (ICS-6), Forschungszentrum Jülich, Germany.|Institut für Physikalische Biologie, Heinrich-Heine-Universität Düsseldorf, Germany.
authors.forename: Bernd W
authors.initials: BW
authors.lastname: Koenig
mesh_terms: D000595:Amino Acid Sequence; D015704:CD4 Antigens / Q000378:metabolism*; D003593:Cytoplasm / Q000378:metabolism*; D004721:Endoplasmic Reticulum / Q000378:metabolism*; D015497:HIV-1 / Q000378:metabolism*; D054298:Human Immunodeficiency Virus Proteins / Q000378:metabolism*; D006801:Humans; D009682:Magnetic Resonance Spectroscopy; D008969:Molecular Sequence Data; D011485:Protein Binding; D017434:Protein Structure, Tertiary; D011993:Recombinant Fusion Proteins / Q000378:metabolism*; D054334:Viral Regulatory and Accessory Proteins / Q000378:metabolism*; D000086082:Viroporin Proteins
chemical_list: D015704:CD4 Antigens; D054298:Human Immunodeficiency Virus Proteins; D011993:Recombinant Fusion Proteins; D054334:Viral Regulatory and Accessory Proteins; C514859:vpu protein, Human immunodeficiency virus 1; D000086082:Viroporin Proteins