Generation of Pseudocontact Shifts in Proteins with Lanthanides Using Small "clickable" Nitrilotriacetic Acid and Iminodiacetic Acid Tags

Choy Theng Loh; Bim Graham; Elwy H. Abdelkader; Kellie L. Tuck; Gottfried Otting

doi:10.1002/chem.201406274

Generation of Pseudocontact Shifts in Proteins with Lanthanides Using Small "clickable" Nitrilotriacetic Acid and Iminodiacetic Acid Tags

Choy Theng Loh, Bim Graham, Elwy H. Abdelkader, Kellie L. Tuck, Gottfried Otting^*

^*Corresponding author for this work

Research School of Chemistry

Research output: Contribution to journal › Article › peer-review

27 Citations (Scopus)

Abstract

Pseudocontact shifts (PCS) induced by paramagnetic lanthanide ions provide unique long-range structural information in nuclear magnetic resonance (NMR) spectra, but the site-specific attachment of lanthanide tags to proteins remains a challenge. Here we incorporated p-azido-phenylalanine (AzF) site-specifically into the proteins ubiquitin and GB1, and ligated the AzF residue with alkyne derivatives of small nitrilotriacetic acid and iminodiacetic acid tags using the Cu^I-catalysed "click" reaction. These tags form lanthanide complexes with no or only a small net charge and produced sizeable PCSs with paramagnetic lanthanide ions in all mutants tested. The PCSs were readily fitted by single magnetic susceptibility anisotropy tensors. Protein precipitation during the click reaction was greatly alleviated by the presence of 150 mM NaCl.

Original language	English
Pages (from-to)	5084-5092
Number of pages	9
Journal	Chemistry - A European Journal
Volume	21
Issue number	13
DOIs	https://doi.org/10.1002/chem.201406274
Publication status	Published - 2015

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10.1002/chem.201406274

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title = "Generation of Pseudocontact Shifts in Proteins with Lanthanides Using Small {"}clickable{"} Nitrilotriacetic Acid and Iminodiacetic Acid Tags",

abstract = "Pseudocontact shifts (PCS) induced by paramagnetic lanthanide ions provide unique long-range structural information in nuclear magnetic resonance (NMR) spectra, but the site-specific attachment of lanthanide tags to proteins remains a challenge. Here we incorporated p-azido-phenylalanine (AzF) site-specifically into the proteins ubiquitin and GB1, and ligated the AzF residue with alkyne derivatives of small nitrilotriacetic acid and iminodiacetic acid tags using the CuI-catalysed {"}click{"} reaction. These tags form lanthanide complexes with no or only a small net charge and produced sizeable PCSs with paramagnetic lanthanide ions in all mutants tested. The PCSs were readily fitted by single magnetic susceptibility anisotropy tensors. Protein precipitation during the click reaction was greatly alleviated by the presence of 150 mM NaCl.",

keywords = "click chemistry, lanthanide-binding tags, NMR spectroscopy, p-azido-L-phenylalanine, pseudocontact shifts",

author = "Loh, \{Choy Theng\} and Bim Graham and Abdelkader, \{Elwy H.\} and Tuck, \{Kellie L.\} and Gottfried Otting",

note = "{\textcopyright} 2015 Wiley-VCH Verlag GmbH \& Co. KGaA.",

year = "2015",

doi = "10.1002/chem.201406274",

language = "English",

volume = "21",

pages = "5084--5092",

journal = "Chemistry - A European Journal",

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T1 - Generation of Pseudocontact Shifts in Proteins with Lanthanides Using Small "clickable" Nitrilotriacetic Acid and Iminodiacetic Acid Tags

AU - Loh, Choy Theng

AU - Graham, Bim

AU - Abdelkader, Elwy H.

AU - Tuck, Kellie L.

AU - Otting, Gottfried

PY - 2015

Y1 - 2015

N2 - Pseudocontact shifts (PCS) induced by paramagnetic lanthanide ions provide unique long-range structural information in nuclear magnetic resonance (NMR) spectra, but the site-specific attachment of lanthanide tags to proteins remains a challenge. Here we incorporated p-azido-phenylalanine (AzF) site-specifically into the proteins ubiquitin and GB1, and ligated the AzF residue with alkyne derivatives of small nitrilotriacetic acid and iminodiacetic acid tags using the CuI-catalysed "click" reaction. These tags form lanthanide complexes with no or only a small net charge and produced sizeable PCSs with paramagnetic lanthanide ions in all mutants tested. The PCSs were readily fitted by single magnetic susceptibility anisotropy tensors. Protein precipitation during the click reaction was greatly alleviated by the presence of 150 mM NaCl.

AB - Pseudocontact shifts (PCS) induced by paramagnetic lanthanide ions provide unique long-range structural information in nuclear magnetic resonance (NMR) spectra, but the site-specific attachment of lanthanide tags to proteins remains a challenge. Here we incorporated p-azido-phenylalanine (AzF) site-specifically into the proteins ubiquitin and GB1, and ligated the AzF residue with alkyne derivatives of small nitrilotriacetic acid and iminodiacetic acid tags using the CuI-catalysed "click" reaction. These tags form lanthanide complexes with no or only a small net charge and produced sizeable PCSs with paramagnetic lanthanide ions in all mutants tested. The PCSs were readily fitted by single magnetic susceptibility anisotropy tensors. Protein precipitation during the click reaction was greatly alleviated by the presence of 150 mM NaCl.

KW - click chemistry

KW - lanthanide-binding tags

KW - NMR spectroscopy

KW - p-azido-L-phenylalanine

KW - pseudocontact shifts

UR - https://www.scopus.com/pages/publications/84924670411

U2 - 10.1002/chem.201406274

DO - 10.1002/chem.201406274

M3 - Article

C2 - 25704668

SN - 0947-6539

VL - 21

SP - 5084

EP - 5092

JO - Chemistry - A European Journal

JF - Chemistry - A European Journal

IS - 13

ER -

Generation of Pseudocontact Shifts in Proteins with Lanthanides Using Small "clickable" Nitrilotriacetic Acid and Iminodiacetic Acid Tags

Abstract

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