Journal: ACS Chemical Biology

Article Title: High Divergence of the Precursor Peptides in Combinatorial Lanthipeptide Biosynthesis

doi: 10.1021/cb500622c

Figure Lengend Snippet: Modification of ProcAt.1 by ProcM. (A) MALDI-ToF-MS analysis of ProcAt.1 that was obtained by coexpression with ProcM and treated with endoproteinase Glu-C (trace i) and subsequently derivatized by NEM (trace ii). (B) Sequence of ProcAt.1 modified by ProcM and treated with Glu-C. The ESI-MS/MS fragmentation pattern for the 3-fold dehydrated species is shown (the MS/MS data is presented in ).

Article Snippet: Restriction endonucleases, DNA polymerases, T4 DNA ligase, and endoproteinase Asp-N and Glu-C were purchased from New England Biolabs.

Techniques: Modification, Sequencing, Tandem Mass Spectroscopy

Journal: ACS Chemical Biology

Article Title: High Divergence of the Precursor Peptides in Combinatorial Lanthipeptide Biosynthesis

doi: 10.1021/cb500622c

Figure Lengend Snippet: Coexpression studies of Cys-lacking peptides with LanMs. (A) MALDI-ToF MS analysis of ProcA4.1 that was obtained by coexpression with ProcM. Also shown is the sequence of the ProcA4.1 core (obtained by TEV cleavage of a ProcA4.1 mutant containing an engineered TEV cleavage site just before the predicted core sequence) and the MS/MS fragmentation pattern for the 3-fold dehydrated species. (B) MALDI-ToF MS analysis of NpnA3 that was obtained by coexpression with NpnM in E. coli . Also shown is the sequence of endoproteinase Glu-C cleaved NpnA3 and the MS/MS fragmentation pattern for the 4-fold dehydrated species. (C) MALDI-ToF MS analysis of NpnA6 obtained similarly to that for NpnA3 in panel B. Also presented is the sequence and MS/MS fragmentation pattern for 3-fold dehydrated NpnA6. The MS/MS data for 3-fold dehydrated ProcA4.1, 4-fold dehydrated NpnA3, and 3-fold dehydrated NpnA6 are shown in , respectively.

Article Snippet: Restriction endonucleases, DNA polymerases, T4 DNA ligase, and endoproteinase Asp-N and Glu-C were purchased from New England Biolabs.

Techniques: Sequencing, Mutagenesis, Tandem Mass Spectroscopy