PXD051183

PXD051183 is an original dataset announced via ProteomeXchange.

Title	A cell-free system for functional studies of small membrane proteins
Description	Numerous small proteins have been discovered across all domains of life, among which many are hydrophobic and predicted to localize to the cell membrane. Based on a few that are well-studied, small membrane proteins are regulators involved in various biological processes, such as cell signaling, nutrient transport, drug resistance, and stress response. However, the function of most identified small membrane proteins remains elusive. Their small size and hydrophobicity make protein production challenging, hindering function discovery. Here, we combined a cell-free system with lipid sponge droplets and synthesized small membrane proteins in vitro. Lipid sponge droplets contain a dense network of lipid bilayers, which accommodates and extracts newly synthesized small membrane proteins from the aqueous surroundings. Using small bacterial membrane proteins MgrB, SafA, and AcrZ as proof of principle, we showed that the in vitro produced membrane proteins were functionally active, for example, modulating the activity of their target kinase as expected. Furthermore, small proteins produced in this system could be used for in vitro co-immunoprecipitations to identify interaction partners. The cell-free system produced small membrane proteins, including one from human, up to micromolar concentrations, indicating its high level of versatility. This work presents a robust alternative approach for producing small membrane proteins, which opens a door to their function discovery in different domains of life.
HostingRepository	PRIDE
AnnounceDate	2025-05-06
AnnouncementXML	Submission_2025-05-06_13:32:36.398.xml
DigitalObjectIdentifier	https://dx.doi.org/10.6019/PXD051183
ReviewLevel	Peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	Supported dataset by repository
PrimarySubmitter	Timo Glatter
SpeciesList	scientific name: Escherichia coli; NCBI TaxID: 562;
ModificationList	No PTMs are included in the dataset
Instrument	Orbitrap Exploris 480

Revision	Datetime	Status	ChangeLog Entry
0	2024-04-03 10:45:48	ID requested
⏵ 1	2025-05-06 13:32:37	announced

10.1016/j.jbc.2024.107850;

10.6019/PXD051183;

Jiang S, Ç, elen G, Glatter T, Niederholtmeyer H, Yuan J, A cell-free system for functional studies of small membrane proteins. J Biol Chem, 300(11):107850(2024) [pubmed]

submitter keyword: Small proteins, protein-protein interaction, in-vitro translation

Timo Glatter
contact affiliation	Max Planck Institute for terrestrial Microbiology Karl-von-Frisch Str. 10 35043 Marburg Germany
contact email	timo.glatter@mpi-marburg.mpg.de
lab head
Timo Glatter
contact affiliation	Max Planck Institute for Terrestrial Microbiology Marburg
contact email	timo.glatter@mpi-marburg.mpg.de
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/2025/05/PXD051183

PRIDE project URI

• PRIDE
  1. PXD051183
     1. Label: PRIDE project
     2. Name: A cell-free system for functional studies of small membrane proteins