Immuno-
precipitation

anti-SA


 homogeneous
F Fo ATP synthase
 with b subunit
 heteroclimers


 solubilize ~I


preps purified


5-10 L KM2
coexpressing
tagged b's


S15 crn 15 % I
SSDS-PAGE I


silver stai n


Figure B-1. Diagram of purification procedures for homogeneous heterodimeric bvs/bhis
 F1Fo ATP synthase complexes. The b subunit dimers represents intact F1Fo
 ATP synthase. Basically, membrane preparations will be solubilized in 0.2%
 tegamineoxide WS-35. The solubilized proteins will then be Ni-resin purified
 in order to remove the homodimeric bvs/bys F1Fo complexes. This will be
 followed by a V5 immunoprecipitation step to remove the homodimeric
 bhis/bhis F1Fo complexes, leaving only the heterodimeric bV5/bhis COmplexes in
 solution. A battery of tests will then be run in order to determine the success
 of the purification procedures and then ultimately activity assays will be
 performed.


Activity
SAssays


S10-18% gradient I
S DS-PAGE I


Western &
Densitornetry