New Lanthanide Chelating Tags for PCS NMR Spectroscopy with Reduction Stable, Rigid Linkers for Fast and Irreversible Conjugation to Proteins

Bioconjugate Chemistry
Thomas MüntenerDaniel Häussinger

Abstract

Lanthanide chelating tags (LCTs) have been used with great success for determining structures and interactions of proteins and other biological macromolecules. Recently LCTs have also been used for in-cell NMR spectroscopy, but the bottleneck especially for demanding applications like pseudocontact shift (PCS) NMR is the sparse availability of suitable tags that allow for site-selective, rigid, irreversible, fast, and quantitative conjugation of chelated paramagnetic lanthanide ions to proteins via reduction stable bonds. We report here several such tags and focus on a new pyridine thiazole derivate of DOTA, that combines high affinity, rigidity, and selectivity with unprecedented tagging properties. The conjugation to the cysteine thiol of the protein results in a reductively stable thioether bond and proceeds virtually quantitatively in less than 30 min at 100 μM protein concentration, ambient temperature, and neutral pH. Upon conjugation of the new tag to two single cysteine mutants of ubiquitin and a single cysteine mutant of human carbonic anhydrase type II (30 kDa) only one stereoisomer is formed (square antiprismatic coordination, Λ(δδδδ)) and large to very large pseudocontact shifts as well as large residual dipolar cou...Continue Reading

References

Jun 24, 2004·Journal of Biomolecular NMR·Takahisa IkegamiChristian Griesinger
Jun 24, 2004·Journal of Biomolecular NMR·Guido PintacudaGottfried Otting
Nov 23, 2006·Chemistry : a European Journal·Monica D VlasieMarcellus Ubbink
Sep 30, 2009·Journal of the American Chemical Society·Daniel HäussingerStephan Grzesiek
Sep 22, 2012·Journal of the American Chemical Society·Wei-Min LiuMarcellus Ubbink
Nov 17, 2012·Chemistry : a European Journal·Yin YangXun-Cheng Su
Apr 12, 2014·Current Opinion in Structural Biology·Mathias A S Hass, Marcellus Ubbink
Apr 17, 2014·Chemistry : a European Journal·Wei-Min LiuMarcellus Ubbink
Nov 25, 2014·Journal of Biomolecular NMR·Mauro RinaldelliClaudio Luchinat
Feb 10, 2016·Dalton Transactions : an International Journal of Inorganic Chemistry·Ana Christina L OpinaOlga Vasalatiy
Jun 3, 2016·Chemical Communications : Chem Comm·Michael D LeeBim Graham
Jul 6, 2016·The Journal of Physical Chemistry Letters·Thomas MüntenerFrançois-Xavier Theillet
Jul 30, 2016·Chemical Communications : Chem Comm·Bin-Bin PanXun-Cheng Su
Oct 7, 2016·Physical Chemistry Chemical Physics : PCCP·Elizaveta A Suturina, Ilya Kuprov
Mar 12, 2017·Progress in Nuclear Magnetic Resonance Spectroscopy·Christoph Nitsche, Gottfried Otting
Mar 28, 2018·Solid State Nuclear Magnetic Resonance·Sabu VargheseStephen Wimperis

❮ Previous
Next ❯

Citations

Jul 18, 2019·Angewandte Chemie·Qing MiaoMarcellus Ubbink
Aug 16, 2019·Chemical Communications : Chem Comm·Daniel Joss, Daniel Häussinger
Jun 12, 2020·Journal of Biomolecular NMR·Charlotte A SoftleyMichael Sattler
May 22, 2019·International Journal of Molecular Sciences·Teppei IkeyaYutaka Ito
Jun 12, 2019·Chemical Science·Kaspar ZimmermannDaniel Häussinger
Jul 6, 2020·Journal of Biomolecular NMR·Thomas MüntenerSebastian Hiller
Nov 18, 2020·Chemistry : a European Journal·Qing MiaoMarcellus Ubbink
Nov 30, 2019·Progress in Nuclear Magnetic Resonance Spectroscopy·Daniel Joss, Daniel Häussinger
Jun 25, 2021·Chemical Science·Zhaofei ChaiConggang Li
Aug 3, 2021·Journal of Biomolecular NMR·Stefano CucuzzaOliver Zerbe
Nov 13, 2021·Chemical Reviews·Sai Chaitanya ChiliveriAd Bax
Jan 11, 2022·Chemical Reviews·Thomas MüntenerSebastian Hiller
Jan 28, 2022·Chemical Reviews·Qing MiaoMarcellus Ubbink

❮ Previous
Next ❯

Related Concepts

Trending Feeds

COVID-19

Coronaviruses encompass a large family of viruses that cause the common cold as well as more serious diseases, such as the ongoing outbreak of coronavirus disease 2019 (COVID-19; formally known as 2019-nCoV). Coronaviruses can spread from animals to humans; symptoms include fever, cough, shortness of breath, and breathing difficulties; in more severe cases, infection can lead to death. This feed covers recent research on COVID-19.

Blastomycosis

Blastomycosis fungal infections spread through inhaling Blastomyces dermatitidis spores. Discover the latest research on blastomycosis fungal infections here.

Nuclear Pore Complex in ALS/FTD

Alterations in nucleocytoplasmic transport, controlled by the nuclear pore complex, may be involved in the pathomechanism underlying multiple neurodegenerative diseases including Amyotrophic Lateral Sclerosis and Frontotemporal Dementia. Here is the latest research on the nuclear pore complex in ALS and FTD.

Applications of Molecular Barcoding

The concept of molecular barcoding is that each original DNA or RNA molecule is attached to a unique sequence barcode. Sequence reads having different barcodes represent different original molecules, while sequence reads having the same barcode are results of PCR duplication from one original molecule. Discover the latest research on molecular barcoding here.

Chronic Fatigue Syndrome

Chronic fatigue syndrome is a disease characterized by unexplained disabling fatigue; the pathology of which is incompletely understood. Discover the latest research on chronic fatigue syndrome here.

Evolution of Pluripotency

Pluripotency refers to the ability of a cell to develop into three primary germ cell layers of the embryo. This feed focuses on the mechanisms that underlie the evolution of pluripotency. Here is the latest research.

Position Effect Variegation

Position Effect Variagation occurs when a gene is inactivated due to its positioning near heterochromatic regions within a chromosome. Discover the latest research on Position Effect Variagation here.

STING Receptor Agonists

Stimulator of IFN genes (STING) are a group of transmembrane proteins that are involved in the induction of type I interferon that is important in the innate immune response. The stimulation of STING has been an active area of research in the treatment of cancer and infectious diseases. Here is the latest research on STING receptor agonists.

Microbicide

Microbicides are products that can be applied to vaginal or rectal mucosal surfaces with the goal of preventing, or at least significantly reducing, the transmission of sexually transmitted infections. Here is the latest research on microbicides.