A novel method for detection of selenomethionine incorporation in protein crystals via Raman microscopy(710 views) Vergara A, Merlino A, Pizzo E, D'Alessio G, Mazzarella L
Dipartimento di Chimica, Università degli Studi di Napoli 'Federico II', Complesso Monte S. Angelo, Via Cinthia, I-80126 Napoli, Italy
Istituto di Biostrutture e Bioimmagini, CNR, Via Mezzocannone 6, I-80134 Napoli, Italy
Consorzio BioTekNet, Università di Napoli Federico II, Napoli, Italy
Dipartimento di Biologia Strutturale e Funzionale, Università di Napoli Federico II, Napoli, Italy
References: Not available.
A novel method for detection of selenomethionine incorporation in protein crystals via Raman microscopy
Multiwavelength anomalous dispersion (MAD) is the most widespread approach in structural biology for the determination of the crystal structure of a novel protein. Mass spectrometry is currently used to evaluate the selenomethionine (SeMet) content in solution, but excluding fluorescence spectroscopy at the absorption edge, no other routine method to check the SeMet incorporation and storage in the crystal state is yet available. Raman microscopy is an increasingly popular tool in physical biochemistry, with applications ranging from studies of ligand binding to secondary-structure analysis. Here, a novel methodological development is presented for the analysis via Raman microscopy of SeMetlabelled protein crystals to be used for MAD crystallography. The method is described and supported by validation and application to two novel proteins (a beta gamma-crystallin-like protein and a DNA-binding protein). Markers of the SeMet residues are in the range 570-600 cm(-1), where proteins do not usually show Raman bands.
A novel method for detection of selenomethionine incorporation in protein crystals via Raman microscopy
Petraglia F, Singh AA, Carafa V, Nebbioso A, Conte M, Scisciola L, Valente S, Baldi A, Mandoli A, Petrizzi VB, Ingenito C, De Falco S, Cicatiello V, Apicella I, Janssen-megens EM, Kim B, Yi G, Logie C, Heath S, Ruvo M, Wierenga ATJ, Flicek P, Yaspo ML, Della Valle V, Bernard O, Tomassi S, Novellino E, Feoli A, Sbardella G, Gut I, Vellenga E, Stunnenberg HG, Mai A, Martens JHA, Altucci L * Combined HAT/EZH2 modulation leads to cancer-selective cell death(455 views) Oncotarget (ISSN: 1949-2553electronic, 1949-2553linking), 2018 May 22; 9(39): 25630-25646. Impact Factor:5.008 ViewExport to BibTeXExport to EndNote