TABLE 1

Cysteine scanning mutagenesis of transmembrane domain

7 hASBT cDNA encoding the C270A mutation were used as the template to generate each of the 22 cysteine double mutants (i.e. Cys270Ala/Phe287Cys using oligonucleotide site-directed mutagenesis). C270A proves resistant to MTS modification and was hence used as the template for all mutations.


No.

Amino Acid Mutationa

Codon Change
1 Phe287Cys TTC→TGC
2 Thr288Cys ACC→TGC
3 Phe289Cys TTC→TGC
4 Pro290Cys CCG→TGC
5 Leu291Cys CTC→TGC
6 Ile292Cys ATC→TGC
7 Tyr293Cys TAC→TGC
8 Ser294Cys AGC→TGC
9 Ile295Cys ATT→TGT
10 Phe296Cys TTC→TGC
11 Glu297Cys CAG→TGC
12 Leu298Cys CTC→TGC
13 Ala299Cys GCC→TGC
14 Phe300Cys TTT→TGT
15 Ala301Cys GCC→TGC
16 Ala302Cys GCA→TGT
17 Ile303Cys ATA→TGT
18 Phe304Cys TTC→TGC
19 Leu305Cys TTA→TGT
20 Gly306Cys GGA→TGT
21 Phe307Cys TTT→TGT
22
Tyr308Cys
TAT→TGT
  • a Represents the position of the mutation of a particular amino acid to a cysteine.