Synthesis of rhamnosylated arginine glycopeptides and determination of the glycosidic linkage in bacterial elongation factor P.
Siyao WangLeo CorciliusPhillip P SharpAndrei RajkovicMichael IbbaBenjamin L ParkerRichard J PaynePublished in: Chemical science (2016)
A new class of N-linked protein glycosylation - arginine rhamnosylation - has recently been discovered as a critical modification for the function of bacterial elongation factor P (EF-P). Herein, we describe the synthesis of suitably protected α- and β-rhamnosylated arginine amino acid "cassettes" that can be directly installed into rhamnosylated peptides. Preparation of a proteolytic fragment of Pseudomonas aeruginosa EF-P bearing both α- and β-rhamnosylated arginine enabled the unequivocal determination of the native glycosidic linkage to be α through 2D NMR and nano-UHPLC-tandem mass spectrometry studies.
Keyphrases
- amino acid
- solid phase extraction
- tandem mass spectrometry
- nitric oxide
- ultra high performance liquid chromatography
- molecularly imprinted
- high performance liquid chromatography
- simultaneous determination
- pseudomonas aeruginosa
- liquid chromatography
- gas chromatography
- high resolution
- genome wide
- magnetic resonance
- high resolution mass spectrometry
- ms ms
- cystic fibrosis
- mass spectrometry
- gene expression
- biofilm formation
- acinetobacter baumannii
- small molecule
- solid state
- protein protein