ArticleMolecular biology of the cell2009
Specific biarsenical labeling of cell surface proteins allows fluorescent- and biotin-tagging of amyloid precursor protein and prion proteins.
Article in Molecular biology of the cell, 2009. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 24 papers.
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Who cites it
24 citing papers in PubMed, 51 citations in OpenAlex.
- Fluorescent non-canonical amino acid as a site-specific conformational probe of prion formation.bioRxiv : the preprint server for biology · 2026Article
- Prion Protein Endoproteolysis: Cleavage Sites, Mechanisms and Connections to Prion Disease.Journal of neurochemistry · 2025Review
- PrP Knockout Cells Expressing Transmembrane PrP Resist Prion Infection.Journal of virology · 2017Article
- Synthesis of Arylazide- and Diazirine-Containing CrAsH-EDT2 Photoaffinity Probes.Archiv der Pharmazie · 2016Article
- Prion protein "gamma-cleavage": characterizing a novel endoproteolytic processing event.Cellular and molecular life sciences : CMLS · 2016Article
- Arf6 controls beta-amyloid production by regulating macropinocytosis of the Amyloid Precursor Protein to lysosomes.Molecular brain · 2015Article
- Glycosylphosphatidylinositol anchoring directs the assembly of Sup35NM protein into non-fibrillar, membrane-bound aggregates.The Journal of biological chemistry · 2014Article
- Live imaging of prions reveals nascent PrPSc in cell-surface, raft-associated amyloid strings and webs.The Journal of cell biology · 2014Article
- Analysis of borna disease virus trafficking in live infected cells by using a virus encoding a tetracysteine-tagged p protein.Journal of virology · 2013Article
- Effects of FlAsH/tetracysteine (TC) Tag on PrP proteolysis and PrPres formation by TC-scanning.Chembiochem : a European journal of chemical biology · 2013Article
- Critical significance of the region between Helix 1 and 2 for efficient dominant-negative inhibition by conversion-incompetent prion protein.PLoS pathogens · 2013Article
- Integrity of helix 2-helix 3 domain of the PrP protein is not mandatory for prion replication.The Journal of biological chemistry · 2012Article
- Cellular prion protein regulates its own α-cleavage through ADAM8 in skeletal muscle.The Journal of biological chemistry · 2012Article
- A naturally occurring C-terminal fragment of the prion protein (PrP) delays disease and acts as a dominant-negative inhibitor of PrPSc formation.The Journal of biological chemistry · 2011Article
- Surveying protein structure and function using bis-arsenical small molecules.Accounts of chemical research · 2011Article
- Article
- Variably protease-sensitive prionopathy: a new sporadic disease of the prion protein.Annals of neurology · 2010Article
- Endogenous proteolytic cleavage of disease-associated prion protein to produce C2 fragments is strongly cell- and tissue-dependent.The Journal of biological chemistry · 2010Article
- Article
- Analysis of protein levels of 24 cytokines in scrapie agent-infected brain and glial cell cultures from mice differing in prion protein expression levels.Journal of virology · 2009Article
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Authors and funding
6 authors at 2 institutions in 1 country.
Funding
Abstract
Fluorescent tagging is a powerful tool for imaging proteins in living cells. However, the steric effects imposed by fluorescent tags impair the behavior of many proteins. Here, we report a novel technique, Instant with DTT, EDT, And Low temperature (IDEAL)-labeling, for rapid and specific FlAsH-labeling of tetracysteine-tagged cell surface proteins by using prion protein (PrP) and amyloid precursor protein (APP) as models. In prion-infected cells, FlAsH-labeled tetracysteine-tagged PrP converted from the normal isoform (PrPsen) to the disease-associated isoform (PrPres), suggesting minimal steric effects of the tag. Pulse-chase analysis of PrP and APP by fluorescent gel imaging demonstrated the utility of IDEAL labeling in investigating protein metabolism by identifying an as-yet-unrecognized C-terminal fragment (C3) of PrPsen and by characterizing the kinetics of PrPres and APP metabolism. C3 generation and N-terminal truncation of PrPres were inhibited by the anti-prion compound E64, a cysteine protease inhibitor. Surprisingly, E64 did not inhibit the synthesis of new PrPres, providing insight into the mechanism by which E64 reduces steady-state PrPres levels in prion-infected cells. To expand the versatility of tetracysteine tagging, we created new Alexa Fluor- and biotin-conjugated tetracysteine-binding molecules that were applied to imaging PrP endocytosis and ultrastructural localization. IDEAL-labeling extends the use of biarsenical derivatives to extracellular proteins and beyond microscopic imaging.
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