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Site-Directed Nitroxide Spin Labeling of Biopolymers

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Structural Information from Spin-Labels and Intrinsic Paramagnetic Centres in the Biosciences

Part of the book series: Structure and Bonding ((STRUCTURE,volume 152))

Abstract

The application of electron paramagnetic resonance (EPR) spectroscopy to the study of biopolymer structure and dynamics has seen rapid growth in the last decade. In addition to advances in instrumentation – in particular, the development of high-field spectrometers and pulsed-EPR methods – spin-labeling techniques have evolved. Nitroxide spin labels can now routinely be incorporated at selected sites to interrogate how structure and dynamics at specific locations relate to biopolymer function. Furthermore, spin labels with improved properties have emerged, in particular, rigid labels that yield more accurate distance measurements, give information about orientation, and faithfully report site-specific dynamics. This review recounts how the three main approaches for site-directed spin labeling of biopolymers, namely, postsynthetic labeling, labeling during biopolymer synthesis, and noncovalent labeling, have been used to label proteins as well as nucleic acids.

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Abbreviations

2AP:

2-Aminopurine

4-SU:

4-Thiouridine

ADP:

Adenosine diphosphate

AMP:

Adenosine monophosphate

Arg:

Arginine

ATP:

Adenosine triphosphate

BSA:

Bovine serum albumin

CoA:

Coenzyme A

CW:

Continuous wave

Cys-tRNA:

Cysteine tRNA

dA:

Deoxyadenosine

DEER:

Double electron–electron resonance

DNA:

Deoxyribonucleic acid

DQC:

Double-quantum coherence

EPR:

Electron paramagnetic resonance

Fmoc:

9-Fluorenylmethyloxycarbonyl

Glu-tRNA:

Glutamyl tRNA

GMPS:

Guanosine monophosphorothioate

HPLC:

High-performance liquid chromatography

mRNA:

Messenger ribonucleic acid

MTS:

1-Oxyl-2,2,5,5-tetramethylpyrrolinyl-3-methyl)-methanethiosulfonate

NAD+ :

Nicotinamide adenine dinucleotide

NC-SDSL:

Noncovalent and site-directed spin labeling

NMR:

Nuclear magnetic resonance

PELDOR:

Pulsed electron–electron double resonance

Phe-tRNA:

Phenylalanine tRNA

POAC:

2,2,5,5-Tetramethylpyrrolidine-N-oxyl-3-amino-4-carboxylic acid

RNA:

Ribonucleic acid

SDSL:

Site-directed spin labeling

Ser:

Serine

SPPS:

Solid-phase peptide synthesis

TAR RNA:

Trans-activation responsive RNA

TEMPO:

2,2,6,6-Tetramethylpiperidine-1-oxyl

TOAC:

2,2,6,6-Tetramethylpiperidine-1-oxyl-4-amino-4-carboxylic acid

TPA:

2,2,5,5-Tetramethyl-pyrrolin-1-yloxy-3-acetylene

tRNA:

Transfer ribonucleic acid

Tyr:

Tyrosine

UDP:

Uridine diphosphate

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  1. Science Institute, University of Iceland, Dunhaga 3, 107, Reykjavik, Iceland

    Sandip A. Shelke & Snorri Th. Sigurdsson

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  1. Sandip A. Shelke
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  2. Snorri Th. Sigurdsson
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Correspondence to Snorri Th. Sigurdsson .

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  1. Inorganic Chemistry Laboratory, University of Oxford, Oxford, United Kingdom

    Christiane R. Timmel

  2. Inorganic Chemistry Laboratory, and Centre for Advanced Imaging, University of Oxford, United Kingdom, and University of Queensland, St. Lucia, Queensland, Australia

    Jeffrey R. Harmer

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Shelke, S.A., Sigurdsson, S.T. (2011). Site-Directed Nitroxide Spin Labeling of Biopolymers. In: Timmel, C., Harmer, J. (eds) Structural Information from Spin-Labels and Intrinsic Paramagnetic Centres in the Biosciences. Structure and Bonding, vol 152. Springer, Berlin, Heidelberg. https://doi.org/10.1007/430_2011_62

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