Nef protein of human immunodeficiency virus type 1 binds its own myristoylated N-terminus.
HIV-1 Nef is a small protein (approx. 25 kDa) that is posttranslationally modified by myristoylation. To explain its complex activities, a 'Nef-cycle' is discussed, which postulates different molecular conformations of Nef. Using recombinant full-length non-myristoylated Nef and synthetic peptides, we demonstrate by fluorescence titration experiments that a peptide representing the myristoylated N-terminus of Nef is specifically bound by Nef. A non-myristoylated N-terminal fragment of Nef or a myristoylated control peptide does not bind to Nef. These results are the first direct experimental evidence of the existence of a myristate-binding pocket in Nef, a prerequisite of the postulated 'closed' Nef conformation.
Hoffmann
Silke
S
Institut für Physikalische Biologie and BMFZ, Heinrich-Heine-Universität, D-40225 Düsseldorf, Germany.
Jonas
Esther
E
König
Simone
S
Preusser-Kunze
Andrea
A
Willbold
Dieter
D
D002151:Calorimetry / Q000379:methods; D015693:Gene Products, nef / Q000737:chemistry*; D006801:Humans; D019814:Myristic Acid / Q000737:chemistry*; D010455:Peptides / Q000737:chemistry; D011994:Recombinant Proteins / Q000737:chemistry; D013050:Spectrometry, Fluorescence; D054311:nef Gene Products, Human Immunodeficiency Virus
D015693:Gene Products, nef; D010455:Peptides; D011994:Recombinant Proteins; D054311:nef Gene Products, Human Immunodeficiency Virus; D019814:Myristic Acid