A microscale protein NMR sample screening pipeline.

Journal of Biomolecular NMR
Paolo RossiGaetano T Montelione

Abstract

As part of efforts to develop improved methods for NMR protein sample preparation and structure determination, the Northeast Structural Genomics Consortium (NESG) has implemented an NMR screening pipeline for protein target selection, construct optimization, and buffer optimization, incorporating efficient microscale NMR screening of proteins using a micro-cryoprobe. The process is feasible because the newest generation probe requires only small amounts of protein, typically 30-200 microg in 8-35 microl volume. Extensive automation has been made possible by the combination of database tools, mechanization of key process steps, and the use of a micro-cryoprobe that gives excellent data while requiring little optimization and manual setup. In this perspective, we describe the overall process used by the NESG for screening NMR samples as part of a sample optimization process, assessing optimal construct design and solution conditions, as well as for determining protein rotational correlation times in order to assess protein oligomerization states. Database infrastructure has been developed to allow for flexible implementation of new screening protocols and harvesting of the resulting output. The NESG micro NMR screening pipeline h...Continue Reading

References

Nov 5, 1999·Journal of Molecular Biology·P E Wright, H J Dyson
Jul 27, 2001·Nature Structural Biology·K W Plaxco, M Gross
May 23, 2002·Biochemistry·A Keith DunkerZoran Obradović
Sep 7, 2002·Journal of Molecular Biology·Jinfeng LiuBurkhard Rost
Oct 17, 2002·Journal of Molecular Biology·Lilia M IakouchevaA Keith Dunker
Mar 8, 2003·Analytical Chemistry·Masaya KakutaJonathan V Sweedler
May 29, 2003·Nucleic Acids Research·Chern-Sing GohMark Gerstein
Jun 26, 2003·Nucleic Acids Research·Rune LindingToby J Gibson
Nov 8, 2003·Structure·Rune LindingRobert B Russell
Jan 13, 2004·Proceedings of the National Academy of Sciences of the United States of America·Dennis PantazatosVirgil L Woods
May 6, 2004·Journal of the American Chemical Society·Wolfgang PetiMark O'Neil-Johnson
Jul 21, 2004·Journal of Structural and Functional Genomics·John K EverettGaetano T Montelione
Nov 24, 2004·Protein Science : a Publication of the Protein Society·Glen SpraggonScott A Lesley
Jan 20, 2005·Bioinformatics·Karen Coeytaux, Anne Poupon
Apr 6, 2005·Methods in Enzymology·Thomas B ActonGaetano T Montelione
Aug 23, 2005·Gait & Posture·Gabor BartonSteve Attfield
Sep 29, 2005·Journal of Magnetic Resonance·Donghan LeeKurt Wüthrich
Nov 12, 2005·Journal of Structural and Functional Genomics·Wolfgang PetiKurt Wüthrich
Apr 19, 2006·BMC Bioinformatics·Kang PengZoran Obradovic
Aug 1, 2006·Journal of the American Society for Mass Spectrometry·S Walter Englander
Oct 6, 2006·Bioinformatics·Oxana V GalzitskayaMichail Yu Lobanov
May 15, 2007·Nature Methods·James M AraminiGaetano T Montelione
May 16, 2008·Journal of the American Chemical Society·Qinghai ZhangKurt Wüthrich
Oct 27, 2009·Journal of Structural and Functional Genomics·Lili MaoMasayori Inouye

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Citations

Apr 25, 2012·Journal of Structural and Functional Genomics·Lida K GiffordPaul D Adams
May 31, 2011·The Journal of Biological Chemistry·James M AraminiGaetano T Montelione
Jul 28, 2013·Science·Ayelet VoskoboynikIrving L Weissman
Aug 22, 2015·Protein Science : a Publication of the Protein Society·John K EverettGaetano T Montelione
Mar 5, 2016·Journal of Structural and Functional Genomics·Marek GrabowskiWladek Minor
Aug 22, 2013·Protein Science : a Publication of the Protein Society·Amy M RuschakTomasz L Religa
Jan 21, 2014·Protein Science : a Publication of the Protein Society·Agnieszka A KendrickElan Z Eisenmesser
Aug 5, 2010·Journal of Structural Biology·Anding HuangRolf Boelens
May 15, 2015·Nucleic Acids Research·Sriram AiyerMonica J Roth
Jul 9, 2016·Biomolecular NMR Assignments·Shuangli LiYunhuang Yang
Dec 11, 2017·Biomolecular NMR Assignments·Fabien KiekenTom Lenaerts
Sep 28, 2017·Chembiochem : a European Journal of Chemical Biology·Florian LehnerRobert Silvers
Jan 15, 2010·Protein Science : a Publication of the Protein Society·Davin R JensenBrian F Volkman
Sep 9, 2020·PLoS Biology·Dmitri TolkatchevAlla S Kostyukova
Aug 11, 2019·International Journal of Molecular Sciences·André F FaustinoIvo C Martins
Oct 28, 2019·Acta Biochimica Et Biophysica Sinica·Chenyun GuoDonghai Lin
Apr 22, 2020·Scientific Reports·Benjamin R DudenhoefferStefan H Knauer
Jul 4, 2020·Journal of Biomolecular NMR·Reto HorstJane M Withka
Sep 29, 2018·Journal of Biological Inorganic Chemistry : JBIC : a Publication of the Society of Biological Inorganic Chemistry·Chris A E M SpronkStefano Ciurli
Jun 24, 2020·Proceedings of the National Academy of Sciences of the United States of America·Siva L S VelivelliDilip M Shah
Feb 6, 2019·Proceedings of the National Academy of Sciences of the United States of America·Nina RipinFrédéric H-T Allain
Jan 22, 2020·Journal of Biomolecular Structure & Dynamics·Sara RagucciAntimo Di Maro

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Methods Mentioned

BETA
NMR
X-ray
PCR
electrophoresis
gel filtration
gel
static light scattering

Software Mentioned

NESG
Prelink
IconNMR
PRED
t1guide
DISEMBL
Primer Prim ’ er
SPiNE
DRIP
NESG SPiNE

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