PXD028512

PXD028512 is an original dataset announced via ProteomeXchange.

Title	Competitive Binding and Molecular Crowding Regulate the Cytoplasmic Interactome of Non-Viral Polymeric Gene Delivery Vectors
Description	In contrast to the processes controlling the complexation, targeting and uptake of polycationic gene delivery vectors, the molecular mechanisms regulating their cytoplasmic dissociation remains poorly understood. Upon cytosolic entry, vectors become exposed to a complex, concentrated mixture of molecules and biomacromolecules. In this report, we characterise the cytoplasmic interactome associated with a polycationic vector based on poly(dimethylaminoethyl methacrylate) (PDMAEMA) and poly(2-methacrylolyloxyethyltrimethylammonium chloride) (PMETAC) brushes. To quantify the contribution of different classes of low molar mass molecules and biomacromolecules to RNA release, we develop a kinetic model based on competitive binding. Our results identify the importance of competition from highly charged biomacromolecules, such as cytosolic RNA, as a primary regulator of RNA release. Importantly, our data indicate the presence of ribosome associated proteins, proteins associated with translation and transcription factors that may underly a broader impact of polycationic vectors on translation. In addition, we bring evidence that molecular crowding modulates competitive binding and demonstrate how the modulation of such interactions, for example via quaternisation or the design of charge-shifting moieties, impacts on the long-term transfection efficiency of polycationic vectors. Understanding the mechanism regulating cytosolic dissociation will enable the improved design of cationic vectors for long term gene release and therapeutic efficacy.
HostingRepository	PRIDE
AnnounceDate	2022-02-16
AnnouncementXML	Submission_2022-02-16_09:44:34.241.xml
DigitalObjectIdentifier	https://dx.doi.org/10.6019/PXD028512
ReviewLevel	Peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	Supported dataset by repository
PrimarySubmitter	Vinothini Rajeeve
SpeciesList	scientific name: Homo sapiens (Human); NCBI TaxID: 9606;
ModificationList	phosphorylated residue; monohydroxylated residue
Instrument	Q Exactive

Revision	Datetime	Status	ChangeLog Entry
0	2021-09-16 02:37:17	ID requested
⏵ 1	2022-02-16 09:44:35	announced

Raynold AAM, Li D, Chang L, Gautrot JE, Competitive binding and molecular crowding regulate the cytoplasmic interactome of non-viral polymeric gene delivery vectors. Nat Commun, 12(1):6445(2021) [pubmed]

submitter keyword: Gene delivery, polycationic vector, RNA delivery, polymer brush, cytosolic interactome, LC-MS/MS, proteomics

Pedro R. Cutillas, PhD
contact affiliation	Pedro R. Cutillas Professor of Cell Signalling & Proteomics Centre for Genomics and Computational Biology Barts Cancer Institute - a Cancer Research UK Centre of Excellence Queen Mary, University of London John Vane Science Centre, Charterhouse Square, London EC1M 6BQ
contact email	p.cutillas@qmul.ac.uk
lab head
Vinothini Rajeeve
contact affiliation	Centre for Haemato Oncology
contact email	v.rajeeve@qmul.ac.uk
dataset submitter

Dataset FTP location NOTE: Most web browsers have now discontinued native support for FTP access within the browser window. But you can usually install another FTP app (we recommend FileZilla) and configure your browser to launch the external application when you click on this FTP link. Or otherwise, launch an app that supports FTP (like FileZilla) and use this address: ftp://ftp.pride.ebi.ac.uk/pride/data/archive/2022/02/PXD028512

PRIDE project URI

• PRIDE
  1. PXD028512
     1. Label: PRIDE project
     2. Name: Competitive Binding and Molecular Crowding Regulate the Cytoplasmic Interactome of Non-Viral Polymeric Gene Delivery Vectors