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. 2009 Apr 15;284(24):16531–16540. doi: 10.1074/jbc.M109.002980

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© 2009 by The American Society for Biochemistry and Molecular Biology, Inc.

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FIGURE 1. — Effects of b subunit engineering on enzyme coupling as measured by ATP-driven energization of inverted membrane vesicles (34). Membranes prepared from strains KM2/pTAM37/pTAM46 (b_his/b_V5) and KM2/pBR322 (Δb) were used as positive and negative controls, respectively. The point where ATP was added is indicated above each graph, and the expressed b subunits are listed on the right. The traces were obtained in triplicate using a Photon Technologies International spectrofluorimeter, and the results shown are individual representative results. A, traces shown are from membranes prepared from strains KM2/pSBC127 (b_V5), KM2/pSBC128 (b_I76C_,V5), KM2/pSBC129 (b_R83C_,V5), KM2/pSBC130 (b_A90C_,V5), and KM2/pSBC131 (b_E97C_,V5). B, traces shown are from membranes prepared from strains KM2/pSBC97 (Tb), KM2/pSBC123 (Tb_A83C), KM2/pSBC124 (Tb_A90C), KM2/pSBC98 (Tb′), KM2/pSBC125 (Tb′_A83C), KM2/pSBC126 (Tb′_A90C), KM2/pSBC97/pSBC98 (Tb/Tb′), KM2/pSBC123/pSBC125 (Tb_A83C/Tb′_A83C), KM2/pSBC123/pSBC126 (Tb_A83C/Tb′_A90C), KM2/pSBC124/pSBC125 (Tb_A90C/Tb′_A83C), and KM2/pSBC124/pSBC126 (Tb_A90C/Tb′_A90C).