Figure 2.

Western analysis of HIV-1 PR mutants. Extracts from bacteria expressing either the wild type or a PR mutant of pART2 were electrophoresed in a polyacrylamide gel under denaturing conditions. The proteins in the gel were transferred to nitrocellulose and analyzed for the presence of processed reverse transcriptase. p120 indicates the position of the unprocessed Pro–Pol protein precursor, and p66 and p51 indicate the positions of the processed forms of HIV-1 reverse transcriptase. Lane 1 shows staining of reverse transcriptase isolated from virus particles. Lane 2 is the extract from a bacterial culture expressing the wild-type Pro–Pol from the pART2 expression vector. Lane 3 is a negative control for processing in which the codon of the catalytic aspartic acid was changed to encode alanine (D25A) in the pART2 expression vector. Lanes 4–29 are examples of PR mutants with changes at each of 11 positions in the flap [Met-46 (M46) through Val-56 (V56)]. The amino acid substituted in each mutant is shown at the top of the lane using the single letter abbreviation for amino acids.