fmol / µg membrane protein (%CV)b

Apparent Ratioc

Conditionsa

OATP1B1

OATP1B3

OATP2B1

1B1 : 1B3 : 2B1

ABC

210.1 ± 4.9 (5.7)

33.0 ± 1.3 (7.7)

39.5 ± 0.9 (5.4)

5.3 : 0.8 : 1

ABC + Gdn

42.9 ± 1.8 (10.2)

4.1 ± 0.1 (5.1)

23.5 ± 0.4 (4.2)

1.8 : 0.2 : 1

ABC + PMAX

299.8 ± 14.9 (12.2)

66.5 ± 3.9 (11.7)

36.7 ± 0.8 (5.5)

8.2 : 1.8 : 1

ABC + DOC

367.2 ± 8.1 (5.4)

137.2 ± 4.7 (6.8)

43.3 ± 1.3 (7.2)

8.4 : 3.2 : 1

aMembrane fractions were initially denatured in 25 mM ABC alone or 25 mM ABC
containing 7 M Gdn, 0.2% PMAX, or 10% DOC prior to alkylation, dilution, and
digestion.
bData represent the mean ± S.E.M. and coefficient of variation (CV%) for
three transitions measured for each peptide by LC-MS/MS for 44 hr digestions
processed in duplicate. Samples were processed from the same initial pellet for
each transfected cell line to rule out variability in starting material.
cThe apparent ratio between cell lines is provided to demonstrate how preparation
conditions have a differential impact on the quantification of different proteins and
thus alter the apparent ratio that is observed between different proteins.
Table 1: Quantification of OATP1B1 (surrogate-1), OATP1B3 (surrogate-2), and OATP2B1 (surrogate-1) peptides in transfected cell line membrane fractions under different sample preparation conditions.