A fluorescence-based assay for the reductase activity of protein disulfide isomerase. (Articolo in rivista)

Type

• Articolo in rivista (Classe)
• Prodotto della ricerca (Classe)

Label

• A fluorescence-based assay for the reductase activity of protein disulfide isomerase. (Articolo in rivista) (literal)

Anno

• 2006-01-01T00:00:00+01:00 (literal)

Http://www.cnr.it/ontology/cnr/pubblicazioni.owl#doi

• 10.1016/j.ab.2005.11.037 (literal)

Alternative label

• Rossella Tomazzolli;a Mauro Dalla Serra;a Giuseppe Bellisola;b Marco Colombatti;b, Graziano Guella;c,* (2006)
  A fluorescence-based assay for the reductase activity of protein disulfide isomerase.
  in Analytical biochemistry (Print)
  (literal)

Http://www.cnr.it/ontology/cnr/pubblicazioni.owl#autori

• Rossella Tomazzolli;a Mauro Dalla Serra;a Giuseppe Bellisola;b Marco Colombatti;b, Graziano Guella;c,* (literal)

Pagina inizio

• 105 (literal)

Pagina fine

• 112 (literal)

Http://www.cnr.it/ontology/cnr/pubblicazioni.owl#numeroVolume

• 350 (literal)

Rivista

• Analytical biochemistry (Rivista)

Http://www.cnr.it/ontology/cnr/pubblicazioni.owl#numeroFascicolo

• 1 (literal)

Note

• ISI Web of Science (WOS) (literal)

Http://www.cnr.it/ontology/cnr/pubblicazioni.owl#affiliazioni

• a CNR-ITC, Institute of Biophysics, Unit at Trento, Via Sommarive 18, 38050 Povo (Trento), Italy b Section of Immunology, Department of Pathology, University of Verona, L.go L.A.Scuro 10, 37134 Verona, Italy c Laboratory of Bioorganic Chemistry, Department of Physics, University of Trento, Via Sommarive 14, 38050 Povo (Trento), Italy (literal)

Titolo

• A fluorescence-based assay for the reductase activity of protein disulfide isomerase. (literal)

Abstract

• We report on a new spectrofluorimetric assay for the measurement of reductase activity of proteins belonging to the superfamily of thioredoxins such as protein disulfide isomerase (PDI). The assay relies on the preparation of a fluorescence-quenched substrate easily accessible in two steps through functional group transformations of the peptide Gly-Cys-Asp. In the first step fluorescein isothiocyanate is linked to the Gly-NH(2) terminus and in the second step the Cys-SH groups are converted into a disulfide bond. Both intermediate and final substrate have been fully characterized by mass spectrometric and nuclear magnetic resonance measurements. Dimethyl sulfoxide is here reported to be a mild oxidizing agent allowing us to obtain in good overall yield the assay substrate in a single synthetic step. A reliable estimation of PDI reductase activity is obtained via the detection of a strong fluorescence enhancement after enzymatic reduction. Moreover, our assay provides further support for the key role played by thioredoxin reductase in enabling disulfide reductase activity of PDI. (literal)

Prodotto di

• Processi di membrana nella comunicazione intra- ed inter-cellulare (MD.P01.001.001) (Modulo)
• Istituto di biofisica (IBF) (Istituto)

Autore CNR

• ROSSELLA TOMAZZOLLI (Unità di personale esterno)
• MAURO DALLA SERRA (Persona)

Incoming links:

Prodotto

• Istituto di biofisica (IBF) (Istituto)
• Processi di membrana nella comunicazione intra- ed inter-cellulare (MD.P01.001.001) (Modulo)

Autore CNR di

• MAURO DALLA SERRA (Persona)
• ROSSELLA TOMAZZOLLI (Unità di personale esterno)

Http://www.cnr.it/ontology/cnr/pubblicazioni.owl#rivistaDi

• Analytical biochemistry (Rivista)