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. Author manuscript; available in PMC: 2009 Aug 11.
Published in final edited form as: Biochemistry. 2008 Sep 4;47(39):10394–10406. doi: 10.1021/bi8010658

FIGURE 3.

FIGURE 3

Factor Xa digest of CRT-His-BAD PLs to assess topology after reconstitution. (A) Representative avidin and (B) polyHis immunoblots of Dd2 PfCRT-hb digested with Factor Xa: lane 1, biotinylated (A) or polyHis (B) molecular mass standards; lane 2, original undigested PL; lanes 3–7, PL digested for 30, 60, 90, 120, and 150 min, respectively. (C) Plot of the amount of full-length PfCRT-hb remaining over time. Image J (NIH) was used to measure the densitometry of the bands corresponding to PfCRT-hb in the blots in panels A and B. The values were converted to the percent of lane 2, and the results for both blots were averaged; ~76% of the initial protein is cleaved, indicating that at least three-fourths of the molecules are oriented with the C-terminal tail on the outside of the lipid.