FIGURE 9.

The strength of hydrophobic interaction with Pal and the crystal structure of mLolB(L68D/E). A, Pal was released from spheroplasts as a complex with LolA and then incubated with wild-type (wt) mLolB(His) or L68E(His) for 1 h at 30 °C. The reaction mixtures were applied to TALON resin and then eluted with a linear gradient of DDM. Fractions (1 ml) were taken and analyzed by SDS-PAGE and immunoblotting with anti-Pal and -LolB antibodies. The left lanes contained 50% amounts of the input samples. The hydrophobic interaction between Pal and LolA was also examined as described under “Experimental Procedures.” B, superimposition of mLolB, mLolB(L68D), and mLolB(L68E) is represented as tube models in gray, cyan, and magenta, respectively. The side chains of the residue at position 68 are shown as sticks. C, the lipoprotein transfer reaction mediated by Lol proteins and the isolated defective mutants. See “Discussion” for details.