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Measurements were repeated three times and carried out on an itc instrument MicroCal, Northampton, MA. Author contributions XR, Conception and design, Acquisition of data, Analysis and interpretation of data, Drafting or revising the article, Contributed unpublished essential data or reagents. This architecture gives Nef an exceptional degree of plasticity, allowing multiple functions to be encoded within a relatively small structure. Mutational analyses have shown that this site is also required for Nef binding, most likely through recognition of the Nef dileucine motif Mattera et al. The structure shows that all of these residues are well ordered in the complex, in contrast to all previous structures of Nef—effector complexes.

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Direct in vitro binding of full-length human immunodeficiency virus type 1 Nef protein to CD4 cytoplasmic domain. The Nef core is directly involved in contacts as well as serves as a aiks to position the central loop.

John Von Neumann and Herman Goldstine. The intramolecular interactions of H4 are with other sections of the central loop, explaining why this helix has not been observed in other structures of Nef. However, the Nef core also seems to have an important role in organizing this segment.

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This information could be used to develop drugs that work by blocking the amino residues on AP-2 that bind to Nef. The Journal of Experimental Medicine. Figure 4 doesn’t really help much because it shows the Nef interaction footprint, but doesn’t show the positions of any residues.

Ardenet al. The Journal of Aije Biology. Nef interface mutants do not downregulate CD4. Adaptors for clathrin coats: The aiks body of work and its structural mapping are summarized in Table 2. Docking of the unlocked AP A modular polycistronic expression system for overexpressing protein complexes in Escherichia coli.

Strikingly, the exposed face of H1 contains residues Trp57 and Leu58 that have been implicated in direct binding to CD4 Grzesiek et al.

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Nearly all of these Nef residues directly contact AP Data from Nef injections into buffer blanks were subtracted from sample data before analysis.

The funders had no role in study design, data collection and interpretation, or the decision to submit the work for publication. E The key Nef diacidic motif Asp and Zike forms intramolecular hydrogen bonds that stabilize the loop conformation.

Petersenet al.

How HIV-1 Nef hijacks the AP-2 clathrin adaptor to downregulate CD4

AhoWeinbergerKernighan. Data shown are representative of three independent experiments. Nef induces CD4 endocytosis: Model building and refinement were carried out using Coot Emsley et al. This allowed Ren et al. MilnerTofte, Harper and MacQueen. AP-2 complex at the molecular level and identified the amino acid residues within c0099 Nef protein that interact with the AP-2 protein.

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Nef-mediated clathrin-coated pit formation.

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The comparatively high affinity of the interaction and the congruence with previous results with the tetrameric construct led us to conclude that this hemicomplex included all the major determinants of the AP Published online Jan Serine phosphorylation-independent downregulation of ai,e CD4 by nef.

SYP, Acquisition of data, Analysis and interpretation of data. Author information Article notes Copyright and License information Disclaimer. Refined solution structure and backbone dynamics of HIV-1 Nef. A large-scale conformational change couples membrane recruitment to cargo binding in the AP2 clathrin adaptor complex.

The structure also reveals that residues of Nef helix H3 iake the core directly contact AP