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. Author manuscript; available in PMC: 2012 Oct 18.
Published in final edited form as: Biochemistry. 2011 Sep 22;50(41):8853–8861. doi: 10.1021/bi200981a

Figure 2.

Figure 2

Catalysis by TbSLS4 HHD triad mutations. A, translation and proteoliposome purification of TbSLS4 and variants with Ala mutations in the HHD triad shown by SDS-PAGE. The positions of molecular mass standards (kDa) are indicated. BioRad stain-free analysis was used to normalize the amount of TbSLS from these preparations to be used in enzyme reactions. B, thin-layer chromatography fractionation of products from reactions containing either NBD-C6-CM (left panel) or NBD-C6-SM (right panel) and ~5 μg of TbSLS4 or variants with the indicated Ala mutations of the HHD triad. The migration positions of SM and CM were determined from mobility standards, while EPC migration was based on comparison to the known second product of TbSLS2.