PXD062870

PXD062870 is an original dataset announced via ProteomeXchange.

Title	Directed evolution of the multicopper oxidase laccase for cell surface proximity labeling and electron microscopy
Description	Enzymes that oxidize aromatic substrates have shown utility in a range of cell-based technologies including live cell proximity labeling (PL) and electron microscopy (EM), but are associated with drawbacks such as the need for toxic H2O2. Here, we explore laccases as a novel enzyme class for PL and EM in mammalian cells. LaccID, generated via 11 rounds of directed evolution from an ancestral fungal laccase, catalyzes the one-electron oxidation of diverse aromatic substrates using O2 instead of toxic H2O2, and exhibits activity selective to the surface plasma membrane of both living and fixed cells. We show that LaccID can be used with mass spectrometry-based proteomics to map the changing surface composition of T cells that engage with tumor cells via antigen-specific T cell receptors. In addition, we use LaccID as a genetically-encodable tag for EM visualization of cell surface features in mammalian cell culture and in the fly brain. Our study paves the way for future cell-based applications of LaccID.
HostingRepository	PRIDE
AnnounceDate	2025-04-12
AnnouncementXML	Submission_2025-04-11_19:03:27.394.xml
DigitalObjectIdentifier
ReviewLevel	Peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	Unsupported dataset by repository
PrimarySubmitter	Keith Rivera
SpeciesList	scientific name: Homo sapiens (Human); NCBI TaxID: 9606;
ModificationList	monohydroxylated residue; deamidated residue; iodoacetamide derivatized residue
Instrument	Orbitrap Exploris 480

Revision	Datetime	Status	ChangeLog Entry
0	2025-04-11 17:34:08	ID requested
⏵ 1	2025-04-11 19:03:28	announced

Dataset with its publication pending

submitter keyword: TMT,Proximity labeling

Steven A. Carr
contact affiliation	Broad Institute
contact email	scarr@broad.mit.edu
lab head
Keith Rivera
contact affiliation	Broad Institute
contact email	krivera@broadinstitute.org
dataset submitter

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PRIDE project URI

• PRIDE
  1. PXD062870
     1. Label: PRIDE project
     2. Name: Directed evolution of the multicopper oxidase laccase for cell surface proximity labeling and electron microscopy