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Rules of Coexistence: RiPP Recognition Elements Evolved to Prevent Pathway Interference through Leader Peptide Discrimination

ABSTRACT: Identification of the precursor peptide post translational modifications by MALDI-TOF MS followed by Mascot search. LppK kinase modifies conserved Ser3 residue in LppA. LppA peptide fused with N-terminal MBP and C-terminal TrxA mass tag (MW 60.1 kDa) co-expressed with LppB harboring N-terminal Strep-tag and LppK. The LppB-LppA complex was purified using affinity chromatography. Protein zones corresponding to LppA (MBP-LppA-TrxA fused protein, MW 61.5 kDa) were subjected to in-gel digestion with trypsin. The eluted tryptic peptides were identified using MALDI-TOF MS. For the leader peptide cleavage site identification, LppA peptide fused with N-terminal MBP and C-terminal TrxA mass tag (MW 60.1 kDa) co-expressed with LppB harboring N-terminal Strep-tag and LppB2. The LppB-LppA complex was purified using affinity chromatography. Protein zones corresponding to LppA (MBP-LppA-TrxA fused protein, MW 61.5 kDa) were subjected to in-gel digestion with trypsin. The eluted tryptic peptides were identified using MALDI-TOF MS.

ORGANISM(S): Escherichia Coli

SUBMITTER: Svetlana Dubiley

PROVIDER: PXD070677 | JPOST Repository | Thu Nov 13 00:00:00 GMT 2025

REPOSITORIES: jPOST

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			Action	DRS
	MBP-LppA-Trx%20%20S30%C3%90%C2%90%20.zip	Other
	MBP-LppA-Trx%20cut.zip	Other
	MBP-LppA-Trx%20full.zip	Other
	MBP-LppA-trxA%20S30T%20.zip	Other
	MBP-LppA-trxA%20S30Y%20.zip	Other

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