Activity of Cysteine substitution mutant enzymes. Alanine scanning of the 11 cysteines present in the active form C211T was performed to generate a collection of C-to-A mutant forms. The list of clones and their mutations are presented in Additional file 2: Table S1. (A) Clones were tested for production of the mutant protein (top gel) and for their lysoPC-acyltransferase activity (bottom) as described in the legend of Figure 4A. The row number (4 to 17) indicates the clone listed (row 4 to 17) in Additional file 2: Table S1. (B) Activity rate measurements were performed as described in the legend of Figure 4B. In addition to LPCAT1 and C211T, rate values for the mutant enzymes with significant acyltransferase activity as shown on panel A, [C211T C443A, row 4], [C211T C(314,457)A, row 5], [C211T C(216,443,514)A, row 6] and [C211T C(216,314,443,501,514)A, row 7], were determined. Protein expression and activity of the form C211T, C501A are presented in Additional file 1: Figure S1. Values are reported relative to the activity rate obtained with LPCAT1 enzyme. The standard deviation of at least 3 different measurements is indicated as error bars.
Soupene and Kuypers BMC Biochemistry 2012 13:8 doi:10.1186/1471-2091-13-8