Sulfhydryl cross-linking of double LmrA cysteine substitution mutants.

<p>Immunoblotted SDS-polyacrylamide 7.5% gels with 10 μg of membrane protein run in each lane. Lanes 1, 2, and 3 contain S361C, K486C single mutants and the S361C / K486C double mutant, and lanes 4, 5, and 6 contain E520C, P383C single mutants and the P383C / E520C double mutant, respectively. Samples were cross-linked at 4°C in the presence of oxidative copper and ATP. Molecular size positions were run in the lane marked ‘S’. The LmrAe monomer in the lane marked ‘W’ has an apparent molecular mass of 66 kDa.