PXD045277

PXD045277 is an original dataset announced via ProteomeXchange.

Title	Broadening the Utility of Farnesyltransferase-Catalyzed Protein Labeling Using Norbornene–Tetrazine Click Chemistry
Description	Bio-orthogonal chemistry has gained widespread use in the study of many biological systems of interest, including protein prenylation. Prenylation is a post-translational modification in which a 15 or 20 carbon isoprenoid chain is transferred onto a cysteine near the C-terminus of a target protein. The three enzymes, protein farnesyltransferase (FTase), geranylgeranyl transferase I (GGTase I), and geranylgeranyl transferase II (GGTase II), that catalyze this process have been shown to exhibit some variability in substrate selection. This trait has been utilized in the past to transfer an array of farnesyl diphosphate analogues with a range of functionality, like an alkyne- containing analogue for copper-catalyzed bioconjugation reactions for example. Reported here is the synthesis of an analogue of the isoprenoid substrate embedded with norbornene functionality (C10NorOPP) for the study of prenylation and development of multifaceted protein-polymer-label conjugates. This analogue undergoes the inverse electron demand Diels Alder reaction with a tetrazine containing tags, allowing for copper-free labeling of proteins in the prenylome. This probe was synthesized in 7 steps with an overall yield of 7%. The use of C10NorOPP for the study of prenylation was explored in metabolic labeling of prenylated proteins in HeLa, COS-7, and astrocyte cells. Furthermore, in HeLa cells, these modified prenylated proteins were identified and quantified using LFQ proteomics, finding 24 enriched prenylated proteins. Additionally, here we utilize the unique chemistry of C10NorOPP in the construction of a multi- protein-polymer conjugate for the targeted labeling of cancer cells. This combines the specificity of the norbornene-tetrazine conjugation with traditional azide-alkyne cycloaddition for the construction of a polymer linked fluorescent trimer increasing cell binding.
HostingRepository	PRIDE
AnnounceDate	2024-05-21
AnnouncementXML	Submission_2024-05-21_08:55:28.769.xml
DigitalObjectIdentifier
ReviewLevel	Peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	Unsupported dataset by repository
PrimarySubmitter	Shelby Auger
SpeciesList	scientific name: Homo sapiens (Human); NCBI TaxID: 9606;
ModificationList	farnesylated residue; geranylgeranylated residue
Instrument	Orbitrap Fusion

Revision	Datetime	Status	ChangeLog Entry
0	2023-09-11 13:26:53	ID requested
⏵ 1	2024-05-21 08:55:29	announced

Auger SA, Venkatachalapathy S, Suazo KFG, Wang Y, Sarkis AW, Bernhagen K, Justyna K, Schaefer JV, Wollack JW, Pl, ü, ckthun A, Li L, Distefano MD, Broadening the Utility of Farnesyltransferase-Catalyzed Protein Labeling Using Norbornene-Tetrazine Click Chemistry. Bioconjug Chem, 35(7):922-933(2024) [pubmed]

10.1021/acs.bioconjchem.4c00072;

submitter keyword: Prenylation, LFQ, Click Chemistry

Mark D.
contact affiliation	University of Minnesota, Chemistry Department
contact email	diste001@umn.edu
lab head
Shelby Auger
contact affiliation	University of Minnesota
contact email	auger054@umn.edu
dataset submitter

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PRIDE project URI

• PRIDE
  1. PXD045277
     1. Label: PRIDE project
     2. Name: Broadening the Utility of Farnesyltransferase-Catalyzed Protein Labeling Using Norbornene–Tetrazine Click Chemistry