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. Author manuscript; available in PMC: 2010 Sep 15.
Published in final edited form as: Biochemistry. 2009 Sep 15;48(36):8528–8539. doi: 10.1021/bi900616w

Table 1.

TCA kinetic constants for C270A, WT, TM3 and TM7 cysteine mutantsa. Bile acid affinity (Km) for all mutant constructs significantly different from controls, p < 0.05. Asterisks omitted for visual clarity.

Construct Taurocholate kinetics
Kt (μM) Jmax (pmol/min/mg prot)
C270A 7.9 ± 1.95 655.6 ± 32.46
WT 11.6 ± 2.39 176.3 ± 7.08
S128C 17.24 ± 4.74 309.9 ± 20.7
L145C 16.51 ±3.77 281.2 ± 15.47
L146C 15.56 ± 3.52 1223 ± 65.60
I147C 22.72 ± 4.93 1454 ± 82.81
F287C 61.52 ± 5.86 1016 ± 32.77
F289C 34.44 ± 7.37 302.3 ± 18.49
L291C 35.44 ± 6.32 1047 ± 53.77
Y293C 72.03 ± 20.83 1364 ± 72.00
a

COS-1 cells transiently transfected as described in “Materials and Methods”. Uptake measured at TCA concentrations ranging from 0–150 μM (measured in triplicate). Kinetic data analyzed using Graphpad Prism software with constants determined via nonlinear least-square fitting procedures.