“…This variant has been found to behave in an equivalent fashion to wild-type in the previous studies (for example [22]). Reactive loop mutants were prepared on the C232S background using aspartic acid-scanning mutagenesis [12] to generate T345D, A347D, G349D, M351D, L353D, A355D, P357D and S359D substitutions (corresponding with RCL positions P 14 , P 12 , P 10 , P 8 , P 6 , P 4 , P 2 and P 1 in subsite notation [28]), yielding in combination the variants P 14 P 12 , P 10 P 8 , P 6 P 4 , P 2 P 1 , P 6 and P 4 . The pcDNA plasmid containing the Z (E342K) α 1 -antitrypsin allele was used as the basis for the mutants in cell culture experiments [29].…”