In vivo incorporation of selenomethionine in proteins using Pseudomonas aeruginosa as expression host: case study—the outer membrane receptor FpvA

The Crystal Structure of the Pyoverdine Outer Membrane Receptor FpvA from Pseudomonas aeruginosa at 3.6Å Resolution

“…The results of the mass spectrometry analysis on [SeMet]FpvA-Pvd are in agreement with the molecular mass of the selenomethioninelabeled mature FpvA. 26 Therefore, the absence of electron density does not result from an N-terminal proteolysis. For all of the data between 20.0 Å and 3.6 Å (except 4.9% excluded for the R free calculation), the R cryst is 25.9% and the R free 27 (calculated using 2090 reflections) is 28.8%.…”

“…26 The analysis of the self rotation function showed the presence of a non-crystallographic 3-fold axis perpendicular to the b-axis and non-crystallographic 2-fold axes suggesting on the basis of the solvent content that three molecules were in the asymmetric unit. Trials to find 11!3 selenium atoms were carried out but unsuccessful.…”

Post‐Translational Backbone Engineering through Selenomethionine‐Mediated Incorporation of Freidinger Lactams

“…[8,9] In addition, the resulting selenonium salt would be ab etter leaving group than the previously studied sulfonium salts,e ncouraging af acile cyclization. As SeM can be incorporated into peptides by routine SPPS [10] and auxotrophic expression, [11][12][13][14][15][16][17][18][19][20] achemoselective transformation of SeM into al actam would facilitate the generation of backbone-modified peptidomimetics from unprotected protein precursors (Figure 1d). Herein, we present amethod that allows for the straightforward incorporation of Freidinger lactams into unprotected peptides and proteins.…”