Single-molecule peptide fingerprinting.
Jetty van GinkelMike FiliusMalwina SzczepaniakPawel TulinskiAnne S MeyerChirlmin JooPublished in: Proceedings of the National Academy of Sciences of the United States of America (2018)
Proteomic analyses provide essential information on molecular pathways of cellular systems and the state of a living organism. Mass spectrometry is currently the first choice for proteomic analysis. However, the requirement for a large amount of sample renders a small-scale proteomics study challenging. Here, we demonstrate a proof of concept of single-molecule FRET-based protein fingerprinting. We harnessed the AAA+ protease ClpXP to scan peptides. By using donor fluorophore-labeled ClpP, we sequentially read out FRET signals from acceptor-labeled amino acids of peptides. The repurposed ClpXP exhibits unidirectional processing with high processivity and has the potential to detect low-abundance proteins. Our technique is a promising approach for sequencing protein substrates using a small amount of sample.
Keyphrases
- single molecule
- amino acid
- mass spectrometry
- living cells
- atomic force microscopy
- pet imaging
- computed tomography
- liquid chromatography
- protein protein
- label free
- high resolution
- health information
- healthcare
- single cell
- gas chromatography
- magnetic resonance imaging
- small molecule
- magnetic resonance
- antibiotic resistance genes
- wastewater treatment
- social media
- capillary electrophoresis
- ms ms
- high speed