PXD046396

PXD046396 is an original dataset announced via ProteomeXchange.

Title	Mass spectrometry distinguishes putrescine, spermidine, and spermine covalently attached to proteins
Description	Bacterial and mammalian cells are rich in putrescine, spermidine and spermine. Polyamines can be incorporated into proteins in vitro. Very few naturally occurring polyaminated proteins have been identified. Bovine albumin and the recombinant universal stress protein from Francisella tularensis were used as models for mass spectrometry analysis of polyaminated proteins. The proteins were covalently bound to putrescine, spermidine, or spermine by the action of carbodiimide or microbial transglutaminase. Tryptic peptides were subjected to liquid chromatography tandem mass spectrometry. Adducts were identified by Protein Prospector software. We describe the search parameters for identifying polyaminated peptides in Protein Prospector and show convincing MMS spectra for adducts with putrescine, spermidine, and spermine. Manual evaluation led us to recognize signature ions for polyamine adducts on Asp, Glu, and Gln. Manual evaluation recognized neutral loss from putrescine, spermidine and spermine during the fragmentation process. Mechanisms for formation of signature ions and neutral loss are presented. Manual evaluation recognized a false positive adduct which had been formed during trypsinolysis by rearrangement of a peptide sequence. Another false positive was a 71 Da added mass on cysteine, which was initially assumed to be putrescine, but was actually a propionamide adduct for a sample extracted from a polyacrylamide gel. The type of information presented in this report serves as a model for identifying naturally occurring polyaminated proteins.
HostingRepository	PRIDE
AnnounceDate	2024-02-28
AnnouncementXML	Submission_2024-02-28_02:36:53.009.xml
DigitalObjectIdentifier
ReviewLevel	Peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	Unsupported dataset by repository
PrimarySubmitter	Lawrence Schopfer
SpeciesList	scientific name: Bos taurus (Bovine); NCBI TaxID: 9913;
ModificationList	amidated residue
Instrument	Orbitrap Fusion Lumos

Revision	Datetime	Status	ChangeLog Entry
0	2023-10-25 11:58:41	ID requested
⏵ 1	2024-02-28 02:36:53	announced

Schopfer LM, Girardo B, Lockridge O, Larson MA, Universal Stress Protein and Bovine Albumin. Biochem Res Int, 2024():7120208(2024) [pubmed]

10.1155/2024/7120208;

submitter keyword: polyamine, universal stress protein, transglutaminase, mass spectrometry, manual evaluation, carbodiimide, albumin

Oksana Lockridge
contact affiliation	Eppley Institute University of Nebraska Medical Center USA
contact email	olockrid@unmc.edu
lab head
Lawrence Schopfer
contact affiliation	University of Nebraska Medical Center
contact email	lmschopf@unmc.edu
dataset submitter

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PRIDE project URI

• PRIDE
  1. PXD046396
     1. Label: PRIDE project
     2. Name: Mass spectrometry distinguishes putrescine, spermidine, and spermine covalently attached to proteins