ACS Chemical Biology
Articles
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(38) Peptides PCP-9 and PCP-10 required a double coupling of 2.5
equiv of monomer 1 for an additional 1 h at RT to achieve complete
incorporation of the analogue. See the Chemical Synthesis section of
(39) Isolated yields of PCP-peptides containing stable analog 1 are
comparable to the yields of the corresponding phosphopeptides. See
Table S1 for phosphopeptide P-5, P-7, and P-9 isolated yields.
(40) The stability of the methylene-bisphosphonate moiety in
trifluoroacetic acid (TFA) peptide cleavage solutions was also
evaluated. See Table S2 for the stability comparison of peptides
PCP-7 and PP-7 in various TFA cleavage solutions.
(41) Meyer, C., Hoeger, B., Temmerman, K., Tatarek-Nossol, M.,
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̈
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̈
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(49) Of the traditionally used carrier proteins, we selected BSA
because its size (∼67 kDa) allowed for facile monitoring of the ligation
reactions by gel electrophoresis, and the maleimide-activated BSA
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(51) Phosphopeptides P-5, P-7, and P-9 were also treated with dye
15, and fluorescence measurements were taken as control experiments.
See Figure S8 for fluorescence ratios for the phosphopeptides
compared to the PCP-peptide and pyrophosphopeptide ratios.
(52) In addition, ligations of the dephosphorylated peptide 7 and PP-
7 were also successful. The similar staining intensities observed for
ligated peptides PCP-7 and PP-7 demonstrated the similarities of the
chemical properties of the pyrophospho and bisphosphonate moieties
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(35) Although compound 1 was clean prior to chromatography, an
ion exchange chromatography step was required to convert the
product from the sodium salt to the free acid, which improved
solubility in DMF for optimal solid-phase incorporation.
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