title: Pupylated proteins in Corynebacterium glutamicum revealed by MudPIT analysis.
abstract: In a manner similar to ubiquitin, the prokaryotic ubiquitin-like protein (Pup) has been shown to target proteins for degradation via the proteasome in mycobacteria. However, not all actinobacteria possessing the Pup protein also contain a proteasome. In this study, we set out to study pupylation in the proteasome-lacking non-pathogenic model organism Corynebacterium glutamicum. A defined pup deletion mutant of C. glutamicum ATCC 13032 grew aerobically as the parent strain in standard glucose minimal medium, indicating that pupylation is dispensable under these conditions. After expression of a Pup derivative carrying an aminoterminal polyhistidine tag in the Δpup mutant and Ni(2+)-chelate affinity chromatography, pupylated proteins were isolated. Multidimensional protein identification technology (MudPIT) and MALDI-TOF-MS/MS of the elution fraction unraveled 55 proteins being pupylated in C. glutamicum and 66 pupylation sites. Similar to mycobacteria, the majority of pupylated proteins are involved in metabolism or translation. Our results define the first pupylome of an actinobacterial species lacking a proteasome, confirming that other fates besides proteasomal degradation are possible for pupylated proteins.
authors.affiliation: IBG-1: Biotechnology, Institute of Bio- and Geosciences, Forschungszentrum Jülich, Jülich, Germany.
authors.forename: Andreas
authors.initials: A
authors.lastname: Küberl
authors.forename: Benjamin
authors.initials: B
authors.lastname: Fränzel
authors.forename: Lothar
authors.initials: L
authors.lastname: Eggeling
authors.forename: Tino
authors.initials: T
authors.lastname: Polen
authors.forename: Dirk Andreas
authors.initials: DA
authors.lastname: Wolters
authors.forename: Michael
authors.initials: M
authors.lastname: Bott
mesh_terms: D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism*; D002853:Chromatography, Liquid; D048230:Corynebacterium glutamicum / Q000235:genetics / Q000378:metabolism*; D030562:Databases, Protein; D050505:Mutant Proteins / Q000378:metabolism*; D009154:Mutation / Q000235:genetics; D046988:Proteasome Endopeptidase Complex / Q000378:metabolism*; D011499:Protein Processing, Post-Translational; D020543:Proteome / Q000378:metabolism*; D040901:Proteomics / Q000379:methods*; D021241:Spectrometry, Mass, Electrospray Ionization; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D014452:Ubiquitins / Q000378:metabolism*
chemical_list: D001426:Bacterial Proteins; D050505:Mutant Proteins; D020543:Proteome; D014452:Ubiquitins; D046988:Proteasome Endopeptidase Complex
keywords: Corynebacterium glutamicum; Microbiology; MudPIT; Post-translational modification; Protein degradation; Pupylation