A new fluorogenic peptide determines proteasome activity in single cells

Silvana A.M. Urru; Pietro Veglianese; Ada De Luigi; Elena Fumagalli; Eugenio Erba; Rodolfo Gonella Diaza; Andrea Carrà; Enrico Davoli; Tiziana Borsello; Gianluigi Forloni; Niccolò Pengo; Enrico Monzani; Paolo Cascio; Simone Cenci; Roberto Sitia; Mario Salmona

doi:10.1021/jm100362x

A new fluorogenic peptide determines proteasome activity in single cells

Silvana A.M. Urru, Pietro Veglianese, Ada De Luigi, Elena Fumagalli, Eugenio Erba, Rodolfo Gonella Diaza, Andrea Carrà, Enrico Davoli, Tiziana Borsello, Gianluigi Forloni, Niccolò Pengo, Enrico Monzani, Paolo Cascio, Simone Cenci, Roberto Sitia, Mario Salmona

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16 Scopus citations

Abstract

The ubiquitin-proteasome system plays a critical role in many diseases, making it an attractive biomarker and therapeutic target. However, the impact of results obtained in vitro using purified proteasome particles or whole cell extracts is limited by the lack of efficient methods to assess proteasome activity in living cells. We have engineered an internally quenched fluorogenic peptide with a proteasome-specific cleavage motif fused to TAT and linked to the fluorophores DABCYL and EDANS. This peptide penetrates cell membranes and is rapidly cleaved by the proteasomal chymotrypsin-like activity, generating a quantitative fluorescent reporter of in vivo proteasome activity as assessed by time-lapse or flow cytometry fluorescence analysis. This reporter is an innovative tool for monitoring proteasomal proteolytic activities in physiological and pathological conditions.

Original language	English (US)
Pages (from-to)	7452-7460
Number of pages	9
Journal	Journal of medicinal chemistry
Volume	53
Issue number	20
DOIs	https://doi.org/10.1021/jm100362x
State	Published - Oct 28 2010
Externally published	Yes

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Urru, S. A. M., Veglianese, P., De Luigi, A., Fumagalli, E., Erba, E., Gonella Diaza, R., Carrà, A., Davoli, E., Borsello, T., Forloni, G., Pengo, N., Monzani, E., Cascio, P., Cenci, S., Sitia, R., & Salmona, M. (2010). A new fluorogenic peptide determines proteasome activity in single cells. Journal of medicinal chemistry, 53(20), 7452-7460. https://doi.org/10.1021/jm100362x

Urru, SAM, Veglianese, P, De Luigi, A, Fumagalli, E, Erba, E, Gonella Diaza, R, Carrà, A, Davoli, E, Borsello, T, Forloni, G, Pengo, N, Monzani, E, Cascio, P, Cenci, S, Sitia, R & Salmona, M 2010, 'A new fluorogenic peptide determines proteasome activity in single cells', Journal of medicinal chemistry, vol. 53, no. 20, pp. 7452-7460. https://doi.org/10.1021/jm100362x

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AU - Gonella Diaza, Rodolfo

AU - Carrà, Andrea

AU - Davoli, Enrico

AU - Borsello, Tiziana

AU - Forloni, Gianluigi

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A new fluorogenic peptide determines proteasome activity in single cells

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