PXD022744-1

PXD022744 is an original dataset announced via ProteomeXchange.

Title	Rapid enzyme-mediated biotinylation for cell surface proteome profiling
Description	Cell surface is the primary site for sensing the extracellular stimuli. The knowledge of the transient changes on surfaceome upon a perturbation is very important as the initial changed proteins could be driving molecules for some phenotype. However, one or multi-step long-time handling procedures are typically required for efficient labeling by current cell-surface labeling strategies partly due to their slow labeling kinetics. In this study, we developed a fast cell-surface labeling strategy, termed as peroxidase-mediated cell-surface labeling (PECSL), enabling efficient and selective labeling of cell-surface proteins within seconds. With a labeling time of 1 min, 2684 proteins, including 1370 cell surface-annotated proteins (cell surface/plasma membrane/extracellular), 734 transmembrane proteins, and 81 CD antigens, were identified by PECSL technology from HeLa cells. By comparison with negative control experiment using quantitative proteomics, the cytoplasmic contaminations from avidin-affinity purification could be reduced and 500 out of the 731 significantly enriched proteins in positive experimental groups (p-value < 0.05, ≥ 2-fold) were cell surface-annotated proteins, including 393 (54%) plasma membrane proteins. Finally, this technology was applied to track the dynamic changes of surfaceomes upon insulin stimulation at two-time points (5 min and 2 h) in HepG2 cells. 32 proteins were found to be significantly regulated by different regulatory mechanisms, including many known insulin-related proteins, like INSR, CTNNB1, TFRC, IGF2R, SORT1. We envision that this technique could be a powerful tool to analyze the transient changes of surfaceomes with a good time resolution and delineate the temporal and spatial regulation of cellular signaling.
HostingRepository	jPOST
AnnounceDate	2021-11-26
AnnouncementXML	Submission_2021-11-25_07:00:05.279.xml
DigitalObjectIdentifier
ReviewLevel	Non peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	Unsupported dataset by repository
PrimarySubmitter	Yanan Li
SpeciesList	scientific name: Homo sapiens (Human); NCBI TaxID: 9606;
ModificationList	S-carboxamidomethyl-L-cysteine; alpha-amino acetylated residue
Instrument	Q Exactive

Revision	Datetime	Status	ChangeLog Entry
0	2020-11-25 18:58:50	ID requested
⏵ 1	2021-11-25 07:00:05	announced
2	2022-09-18 03:51:26	announced	2022-09-18: Updated FTP location.

Dataset with its publication pending

submitter keyword: Cell surface, plasma membrane, proteomics, enrichment, insulin

Yanan Li
lab head
Yanan Li
contact affiliation	Dalian Institute of Chemical Physics
dataset submitter

jPOST dataset URI

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.biosciencedbc.jp/archive/jpostrepos/JPST001019