A Highly Ordered Nitroxide Side Chain for Distance Mapping and Monitoring Slow Structural Fluctuations in Proteins
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Tartalom: http://real.mtak.hu/176765/
Archívum: REAL
Gyűjtemény: Status = Published
Subject = Q Science / természettudomány: QD Chemistry / kémia: QD02 Physical chemistry / fizikai kémia
Type = Article
Cím:
A Highly Ordered Nitroxide Side Chain for Distance Mapping and Monitoring Slow Structural Fluctuations in Proteins
Létrehozó:
Chen, Mengzhen
Kálai, Tamás
Cascio, Duilio
Bridges, Michael D.
Whitelegge, Julian P.
Elgeti, Matthias
Hubbell, Wayne L.
Kiadó:
Springer-Verlag GmbH Austria
Dátum:
2023
Téma:
QD02 Physical chemistry / fizikai kémia
Tartalmi leírás:
Site-directed spin labeling electron paramagnetic resonance (SDSL-EPR) is an established tool for exploring protein structure and dynamics. Although nitroxide side chains attached to a single cysteine via a disulfide linkage are commonly employed in SDSL-EPR, their internal flexibility complicates applications to monitor slow internal motions in proteins and to structure determination by distance mapping. Moreover, the labile disulfide linkage prohibits the use of reducing agents often needed for protein stability. To enable the application of SDSL-EPR to the measurement of slow internal dynamics, new spin labels with hindered internal motion are desired. Here, we introduce a highly ordered nitroxide side chain, designated R9, attached at a single cysteine residue via a non-reducible thioether linkage. The reaction to introduce R9 is highly selective for solvent-exposed cysteine residues. Structures of R9 at two helical sites in T4 Lysozyme were determined by X-ray crystallography and the mobility in helical sequences was characterized by EPR spectral lineshape analysis, Saturation Transfer EPR, and Saturation Recovery EPR. In addition, interspin distance measurements between pairs of R9 residues are reported. Collectively, all data indicate that R9 will be useful for monitoring slow internal structural fluctuations, and applications to distance mapping via dipolar spectroscopy and relaxation enhancement methods are anticipated.
Nyelv:
angol
Típus:
Article
PeerReviewed
info:eu-repo/semantics/article
Formátum:
text
Azonosító:
Chen, Mengzhen and Kálai, Tamás and Cascio, Duilio and Bridges, Michael D. and Whitelegge, Julian P. and Elgeti, Matthias and Hubbell, Wayne L. (2023) A Highly Ordered Nitroxide Side Chain for Distance Mapping and Monitoring Slow Structural Fluctuations in Proteins. APPLIED MAGNETIC RESONANCE. pp. 1-27. ISSN 0937-9347 (print); 1613-7507 (online)
Kapcsolat:
DOI: 10.1007/s00723-023-01618-8
Létrehozó:
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