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nmrlearner 08-22-2010 02:20 PM

The CD4 determinant for downregulation by HIV-1 Nef directly binds to Nef. Mapping of
 
The CD4 determinant for downregulation by HIV-1 Nef directly binds to Nef. Mapping of the Nef binding surface by NMR.

http://www.ncbi.nlm.nih.gov/corehtml...es-acspubs.jpg Related Articles The CD4 determinant for downregulation by HIV-1 Nef directly binds to Nef. Mapping of the Nef binding surface by NMR.

Biochemistry. 1996 Aug 13;35(32):10256-61

Authors: Grzesiek S, Stahl SJ, Wingfield PT, Bax A

Using heteronuclear NMR spectroscopy, we demonstrate that a 13-residue peptide (MS-QIKRLLSEKKT) from the cytoplasmic tail of CD4 binds to Nef protein. This part of CD4 is critical for downregulation of CD4 by HIV-1 Nef [Aiken et al. (1994) Cell 76, 853-864]. We show that a control peptide without the central dileucine does not bind to Nef. The dependence of Nef 1H and 15N amide chemical shifts on peptide concentration indicates that the binding is in the fast chemical exchange limit, with a dissociation constant Kd of approximately 1 mM. The peptide binding site has been mapped onto the previously determined solution structure of HIV-1 Nef [Grzesiek et al. (1996) Nat. Struct. Biol. 3, 340-345] on the basis of peptide-induced chemical shift changes. It comprises amino acids W57, L58, E59, G95, G96, L97, R106, and L110. When Nef is complexed to the SH3 domain of Hck tyrosine protein kinase, the peptide binds to the same site on Nef but with slightly higher affinity (Kd approximately 0.5 mM). This indicates that the binding of CD4 and Hck SH3 to Nef are two compatible and slightly cooperative events.

PMID: 8756680 [PubMed - indexed for MEDLINE]



Source: PubMed


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