Automated Backbone NMR Resonance Assignment of Large Proteins Using Redundant Linking from a Single Simultaneous Acquisition.

Journal of the American Chemical Society
Jan StanekGuido Pintacuda

Abstract

Thanks to magic-angle spinning (MAS) probes with frequencies of 60-100 kHz, the benefit of high-sensitivity 1H detection can now be broadly realized in biomolecular solid-state NMR for the analysis of microcrystalline, sedimented, or lipid-embedded preparations. Nonetheless, performing the assignment of all resonances remains a rate-limiting step in protein structural studies, and even the latest optimized protocols fail to perform this step when the protein size exceeds ∼20 kDa. Here, we leverage the benefits of fast (100 kHz) MAS and high (800 MHz) magnetic fields to design an approach that lifts this limitation. Through the creation, conservation, and acquisition of independent magnetization pathways within a single triple-resonance MAS NMR experiment, a single self-consistent data set can be acquired, providing enhanced sensitivity, reduced vulnerability to machine or sample instabilities, and highly redundant linking that supports fully automated peak picking and resonance assignment. The method, dubbed RAVASSA (redundant assignment via a single simultaneous acquisition), is demonstrated with the assignment of the largest protein to date in the solid state, the 42.5 kDa maltose binding protein, using a single fully protona...Continue Reading

References

Nov 1, 1995·Journal of Biomolecular NMR·F DelaglioA Bax
Mar 1, 1997·Journal of Magnetic Resonance·M Bak, N C Nielsen
Nov 13, 1998·Proceedings of the National Academy of Sciences of the United States of America·M SalzmannK Wüthrich
Dec 1, 2000·Journal of Magnetic Resonance·M BakN C Nielsen
Sep 5, 2001·Journal of Magnetic Resonance·B ReifR G Griffin
Jul 25, 2003·Journal of Biomolecular NMR·Daniel MalmodinMartin Billeter
Dec 18, 2003·Journal of the American Chemical Society·Eric K PaulsonKurt W Zilm
Jan 23, 2010·Journal of the American Chemical Society·Yun HanTatyana Polenova
Sep 17, 2011·Journal of the American Chemical Society·Meaghan E WardVladimir Ladizhansky
Jul 17, 2012·Journal of the American Chemical Society·Elena Schmidt, Peter Güntert
Aug 9, 2012·Journal of Biomolecular NMR·Ivano BertiniEnrico Ravera
May 17, 2013·The Journal of Chemical Physics·T Gopinath, Gianluigi Veglia
May 22, 2013·Journal of Biomolecular NMR·Elena SchmidtPeter Güntert
Aug 8, 2014·Journal of the American Chemical Society·Emeline Barbet-MassinGuido Pintacuda
Jan 9, 2015·PLoS Computational Biology·Markus NiklassonPatrik Lundström
Mar 24, 2015·Journal of Magnetic Resonance·Loren B AndreasGuido Pintacuda
Mar 24, 2015·Journal of Magnetic Resonance·Anja BöckmannBeat H Meier
May 16, 2015·Journal of Biomolecular NMR·ShengQi XiangRasmus Linser
Jun 17, 2015·Journal of Biomolecular NMR·Loren B AndreasGuido Pintacuda
Aug 5, 2016·Proceedings of the National Academy of Sciences of the United States of America·Loren B AndreasGuido Pintacuda
Jul 21, 2017·Journal of the American Chemical Society·Daniela LalliGuido Pintacuda
Dec 14, 2017·Nature Communications·Joren S RetelHartmut Oschkinat
Feb 22, 2018·Journal of Biomolecular NMR·Sebanti Gupta, Robert Tycko
Feb 27, 2018·The Journal of Physical Chemistry Letters·Suresh K VasaRasmus Linser
Jul 6, 2018·Chemical Communications : Chem Comm·Alexander MarchankaTeresa Carlomagno
Jul 25, 2019·Journal of Magnetic Resonance·Ago Samoson
Oct 31, 2019·Nature Communications·Iva PritišanacPeter Güntert

❮ Previous
Next ❯

Citations

Jul 2, 2020·Biomolecular NMR Assignments·Tobias SchubeisLoren B Andreas
Sep 5, 2020·Progress in Nuclear Magnetic Resonance Spectroscopy·Iva PritišanacPeter Güntert
Apr 18, 2021·Biomolecular NMR Assignments·Tobias SchubeisGuido Pintacuda
Apr 30, 2021·Current Opinion in Structural Biology·Marta BonaccorsiGuido Pintacuda
Aug 10, 2021·Nature Reviews. Methods Primers·Bernd ReifMei Hong
Sep 5, 2021·Progress in Nuclear Magnetic Resonance Spectroscopy·Ēriks KupčeTim D W Claridge

❮ 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.