Top-down mass spectrometry on low-resolution instruments: characterization of phosphopantetheinylated carrier domains in polyketide and non-ribosomal biosynthetic pathways.
Mass spectrometry (MS) is an important tool for studying non-ribosomal peptide, polyketide, and fatty acid biosynthesis. Here we describe a new approach using multi-stage tandem MS on a common ion trap instrument to obtain high-resolution measurements of the masses of substrates and intermediates bound to phosphopantetheinylated (holo) carrier proteins. In particular, we report the chemical formulas of 12 diagnostic MS(3) fragments of the phosphopantetheine moiety ejected from holo carrier proteins during MS(2). We demonstrate our method by observing the formation of holo-AcpC, a putative acyl carrier protein from Streptococcus agalactiae.
Meluzzi
Dario
D
Department of Chemistry and Biochemistry, University of California at San Diego, La Jolla, CA 92093-0636, USA.
Zheng
Wei Hao
WH
Hensler
Mary
M
Nizet
Victor
V
Dorrestein
Pieter C
PC
D000213:Acyl Carrier Protein / Q000737:chemistry*; D001426:Bacterial Proteins / Q000737:chemistry; D018942:Macrolides / Q000737:chemistry*; D013058:Mass Spectrometry / Q000295:instrumentation; D008958:Models, Molecular; D015394:Molecular Structure; D010204:Pantetheine / Q000031:analogs & derivatives* / Q000737:chemistry; D010453:Peptide Synthases / Q000737:chemistry*; D015398:Signal Transduction; D013292:Streptococcus agalactiae / Q000737:chemistry / Q000201:enzymology; D017855:Transferases (Other Substituted Phosphate Groups) / Q000737:chemistry
D000213:Acyl Carrier Protein; D001426:Bacterial Proteins; D018942:Macrolides; C082662:phosphopantetheinyl transferase; D010204:Pantetheine; D017855:Transferases (Other Substituted Phosphate Groups); D010453:Peptide Synthases; C104385:non-ribosomal peptide synthase; C003129:4'-phosphopantetheine