AT P





















 +7r-his


 bwt-his
 b+7-his/ bar-vs

 b+7-his/ bwtz,

 O C ~wtbwt-Vhisl wt-V5


 2 min bwt


Figure 2-9. ATP-driven energization of membrane vesicles prepared from uncF(b) gene
 mutants. Cell membrane vesicles were prepared by differential centrifugation
 (see Materials and Methods). Membrane protein (200 Gig) was suspended in 3
 ml of assay buffer (50 mM MOPS, 10 mM MgCl2, pH 7.3). The fluorescent
 dye ACMA was added to a final concentration of 1 CIM, and fluorescence was
 recorded with excitation at 410 nm and emission at 490 nm. ATP was added
 as indicated to a final concentration of 1mM. The samples for each trace have
 been labeled according to the amino acid insertion or deletion and the epitope
 tag, so the strains used as the sources of the samples were as follows: bwt,
 KM2/pKAM14; bwt-vs, KM2/pTAM46; ba7~vs, KM2/pTAM47; bwt-his,
 KM2/pTAM37; b+7-his, KM2/pTAM35; bwt-his/ bwt-vs,
 KM2/pTAM37/pTAM46; b+7-his/bwt-v5, KM2/pTAM35/pTAM46; b 7.~his/bay.~
 vs, KM2/ pTAM3 5/pTAM47.