FIG. 2.

Determination of the region of HIV-2 Gag involved in binding PRP4. (A) Schematic diagram of HIV-2 Gag domain proteins prepared for in vitro binding assays. ³⁵S-labeled HIV-2 Gag truncations were prepared by TNT in vitro translation. (B) In vitro binding of HIV-2 Gag truncations to PRP4 protein. C-terminal protein truncations were analyzed by GST fusion pull-down assay for the ability to bind GST or GST-PRP4 (lanes 1 to 7) immobilized on glutathione-Sepharose beads. Individual Gag subdomains (MA, CA, and NC) were also assayed (lanes 11 to 13) Inputs (lanes 8 to 10 and 14 to 16) were 10% of the amount of the relevant ³⁵S-labeled HIV-2 Gag protein used in each assay. The positions of marker proteins are in kilodaltons.