Journal: The Journal of Biological Chemistry

Article Title: Biochemical characterization of the first step in sulfonolipid biosynthesis in Alistipes finegoldii

doi: 10.1016/j.jbc.2022.102195

Figure Lengend Snippet: Identification and purification of cysteate acyl-ACP transferase. A , TLC separation of 3-keto[ 14 C]capnine (3[ 14 C]KC) arising from SulA activity using cellular lysates from Escherichia coli cells expressing nothing (empty vector control), alfi_1224 (SulA), or alfi_1465 . The thin-layer plate is representative of four independent experiments. B , purified SulA migrates as a single 85 kDa species ( V e = 76.6 ml) based on a calibrated HiLoad 16/600 Superdex 200 column (98.1 kDa theoretical homodimer mass). Left inset , molecular weight calculation using calibration standards described under . Right inset , SDS gel electrophoresis shows SulA purity (∼48 kDa) along with molecular weight standards. C , the sedimentation velocity profile (fringe displacement) was fit to a continuous sedimentation coefficient distribution model c(S). SulA sedimented as a 98.9 kDa homodimer with a sedimentation coefficient of 5.11 S. ACP, acyl carrier protein; SulA, cysteate acyl-ACP transferase.

Article Snippet: E. coli strain BL21(DE3) cells harboring the pET-SulA or pET-SulA(K281A) expression plasmids were grown in 2 l LB medium with 50 μg/ml kanamycin (Gold Biotechnology) at 37 °C and 200 rpm shaking to A 600 = 0.6.

Techniques: Purification, Activity Assay, Expressing, Plasmid Preparation, Molecular Weight, SDS-Gel, Electrophoresis, Sedimentation

Journal: Nature chemical biology

Article Title: Monitoring methionine sulfoxide with stereospecific mechanism-based fluorescent sensors

doi: 10.1038/nchembio.1787

Figure Lengend Snippet: E. coli cells expressing MetSOx ( a ) or MetROx ( b ) were incubated with MetO for ~30 min, and then rinsed to remove the oxidant. Arrows indicate the addition (↓) of MetO and rinsing (↑) the cells for two representative experiments (Assay 1 and Assay 2). MetSOx ( c ) and MetROx ( d ) oxidized fractions calculated from (n = 3), using and , respectively, described in the “ online methods ” section.

Article Snippet: SoluBL21 TM E. coli (Gelantis, San Diego, CA) cells were transformed with an indicated expression vector and grown in Luria-Bertani containing ampicillin (50 μg.ml −1 ) at 37°C.

Techniques: Expressing, Incubation