Figure 1. Schematic display of Nef Interacting (NI) protein design.

(A) Cartoon of interaction motifs within the Nef structure. (B) Protein domains and lipidation signal sequences employed for the design of Nef interacting proteins. The number of different constructs generated and the proposed target sites are shown to the right. (C) Fusion design of four different NI generations. Different recognition domains were assembled in alternating orders and combined with various membrane association motifs to result in different expression constructs, used either for recombinant protein production or in vivo cell expression. (D) Display of the three high affinity binders designed to wrap the interaction surfaces of Nef. (E) SDS PAGE display of selected recombinant Nef and NI proteins used for in vitro binding analyses.