**Acknowledgements**

Dr. John Walter (retired) has assisted with discussions during the writing of this review and was instrumental with establishing the Biomolecular Magnetic Resonance Facility (BMRF), housed at the National Research Council of Canada's Oxford St. Institute.

### **Author details**

Nadine Merkley, Ian Burton, Tobias Karakach and Raymond T. Syvitski\*

\*Address all correspondence to: Ray.Syvitski@nrc-cnrc.gc.ca

Biomolecular Magnetic Resonance Facility (BMRF), National Research Council of Canada, Canada

### **References**


**Author details**

Canada

**References**

1945;9:211-216.

333-356.

Nadine Merkley, Ian Burton, Tobias Karakach and Raymond T. Syvitski\*

Biomolecular Magnetic Resonance Facility (BMRF), National Research Council of Canada,

[1] Zavoisky YK. Relaxation of liquid solutions for perpendicular fields. J Phys USSR

[3] Purcell EM, Torrey HC, Pound RV. Resonance Absorption by Nuclear Magnetic Mo‐

[4] Ernst RR, Anderson WA. Application of fourier transform spectroscopy to magnetic

[5] Gutowsky HS. Nuclear Magnetic Resonance. Annu Rev Phys Chem 1954;5(1):

[6] Williamson MP, Havel TF, Wuthrich K. Solution conformation of proteinase inhibitor IIA from bull seminal plasma by 1H nuclear magnetic resonance and distance geom‐

[7] P Mansfield and PKG. NMR 'diffraction' in solids? Journal of Physics C: Solid State

[8] Jonas J, Gutowsky HS. NMR in Chemistry--An Evergreen. Annu Rev Phys Chem

[9] Gielen M, Willem R, Wrackmeyer B. Advanced applications of NMR to organometal‐

[10] Burton IW, Quilliam MA, Walter JA. Quantitative 1H NMR with external standards: use in preparation of calibration solutions for algal toxins and other natural products.

[12] Cavanagh J, Fairbrother WJ, Palmer III AG, Skelton NJ. Protein NMR Spectroscopy:

[13] Johnson CS. Diffusion ordered nuclear magnetic resonance spectroscopy: principles

[11] Rabi II, Cohen VW. The Nuclear Spin of Sodium. Phys Rev 1933;43(7):582.

Principles and Practice. New York: Academic Press; 1996.

and applications. Prog Nucl Mag Res Sp 1999;34(3-4):203-256.

[2] Bloch F, Hansen WW, Packard M. Nuclear Induction. Phys Rev 1946;69(3-4):127.

\*Address all correspondence to: Ray.Syvitski@nrc-cnrc.gc.ca

82 Using Old Solutions to New Problems - Natural Drug Discovery in the 21st Century

ments in a Solid. Phys Rev 1946;69(1-2):37-38.

resonance. Rev Sci Instrum 1966;37(1):93-102.

lic chemistry. Chichester ; New York: Wiley; 1996.

etry. J Mol Biol 1985;182(2):295-315.

Anal Chem 2005;77(10):3123-3131.

Physics 1973;6(22):L422.

1980;31(1):1-28.


[44] Insect cell media - BioExpress 2000: New results from CIL. ; 05//; Providence, RI; 2005.

[29] Bax A, Ikura M, Kay LE, Torchia DA, Tschudin R. Comparison of different modes of two-dimensional reverse-correlation NMR for the study of proteins. Journal of Mag‐

[30] Bax A, Summers MF. Proton and carbon-13 assignments from sensitivity-enhanced detection of heteronuclear multiple-bond connectivity by 2D multiple quantum

[31] Nyberg NT, Duus JÃ, Sørensen OW. Heteronuclear Two-Bond Correlation: Sup‐ pressing Heteronuclear Three-Bond or Higher NMR Correlations while Enhancing Two-Bond Correlations Even for Vanishing 2JCH. J Am Chem Soc 2005;127(17):

[32] Reynolds WF, Enriquez RG. Choosing the best pulse sequences, acquisition parame‐ ters, postacquisition processing strategies, and probes for natural product structure

[33] Fukushi E. Advanced NMR approaches for a detailed structure analysis of natural

[34] Bross-Walch N, Kuhn T, Moskau D, Zerbe O. Strategies and tools for structure deter‐ mination of natural products using modern methods of NMR spectroscopy. Chem

[35] Claridge TDW, ScienceDirect. High-resolution NMR techniques in organic chemis‐

[37] Noda I, Ozaki Y, Ebooks Corporation. Two-Dimensional Correlation Spectroscopy.

[38] Richards S, Hollerton J, Ebooks Corporation. Essential Practical NMR for Organic

[39] Wuthrich K. Nuclear Magnetic Resonance (NMR) Spectroscopy of Proteins. eLS:

[40] Tugarinov V, Muhandiram R, Ayed A, Kay LE. Four-dimensional NMR spectrosco‐ py of a 723-residue protein: chemical shift assignments and secondary structure of

[41] Sikic K, Tomic S, Carugo O. Systematic comparison of crystal and NMR protein structures deposited in the protein data bank. Open Biochem J 2010;4:83-95.

[42] Shin J, Lee W, Lee W. Structural proteomics by NMR spectroscopy. Expert Rev Pro‐

[43] Acton TB, Xiao R, Anderson S, Aramini J, Buchwald WA, Ciccosanti C, et al. Prepara‐ tion of protein samples for NMR structure, function, and small-molecule screening

malate synthase g. J Am Chem Soc 2002;124(34):10025-10035.

[36] Jacobsen NE, Ebooks Corporation. NMR Spectroscopy Explained. 2007:686.

elucidation by NMR spectroscopy. J Nat Prod 2002;65(2):221-244.

products. Biosci Biotechnol Biochem 2006;70(8):1803-1812.

netic Resonance (1969) 1990;86(2):304-318.

84 Using Old Solutions to New Problems - Natural Drug Discovery in the 21st Century

NMR. J Am Chem Soc 1986;108(8):2093-2094.

6154-6155.

try. 2009:383.

2005:311.

Chemistry. 2010:230.

John Wiley & Sons, Ltd; 2001.

teomics 2008;5(4):589-601.

studies. Methods Enzymol 2011;493:21-60.

Biodivers 2005;2(2):147-177.


[74] Huang YJ, Powers R, Montelione GT. Protein NMR recall, precision, and F-measure scores (RPF scores): structure quality assessment measures based on information re‐ trieval statistics. J Am Chem Soc 2005;127(6):1665-1674.

[58] Muona M, Aranko AS, Raulinaitis V, Iwai H. Segmental isotopic labeling of multidomain and fusion proteins by protein trans-splicing in vivo and in vitro. Nat Protoc

[59] Chen J, Wang J. A segmental labeling strategy for unambiguous determination of do‐ main–domain interactions of large multi-domain proteins. J Biomol NMR 2011;50(4):

[60] Fagerberg L, Jonasson K, von Heijne G, Uhlen M, Berglund L. Prediction of the hu‐

[61] Warschawski DE, Arnold AA, Beaugrand M, Gravel A, Chartrand E, Marcotte I. Choosing membrane mimetics for NMR structural studies of transmembrane pro‐

[62] Qureshi T, Goto NK. Contemporary methods in structure determination of mem‐

[63] Zhu G, Yao X. TROSY-based NMR experiments for NMR studies of large biomole‐

[64] Tzeng S-, Pai M-, Kalodimos CG. NMR studies of large protein systems. Methods in

[65] Kay LE. Solution NMR spectroscopy of supra-molecular systems, why bother? A

[66] Sprangers R, Kay LE. Quantitative dynamics and binding studies of the 20S protea‐

[67] Guntert P. Automated structure determination from NMR spectra. Eur Biophys J

[68] Gronwald W, Kalbitzer HR. Automated structure determination of proteins by NMR

[69] Banci L, Bertini I, Luchinat C, Mori M. NMR in structural proteomics and beyond.

[70] Delaglio F, Wu Z, Bax A. Measurement of Homonuclear Proton Couplings from Reg‐ ular 2D COSY Spectra. Journal of Magnetic Resonance 2001;149(2):276-281.

[71] Shen Y, Delaglio F, Cornilescu G, Bax A. TALOS+: a hybrid method for predicting protein backbone torsion angles from NMR chemical shifts. J Biomol NMR

[72] Jain NU. Use of residual dipolar couplings in structural analysis of protein-ligand complexes by solution NMR spectroscopy. Methods Mol Biol 2009;544:231-252.

[73] Chen K, Tjandra N. The use of residual dipolar coupling in studying proteins by

spectroscopy. Prog Nucl Magn Reson Spectrosc 2004;44(1-2):33-96.

brane proteins by solution NMR. Top Curr Chem 2012;326:123-186.

man membrane proteome. Proteomics 2010;10(6):1141-1149.

teins. Biochim Biophys Acta 2011;1808(8):1957-1974.

86 Using Old Solutions to New Problems - Natural Drug Discovery in the 21st Century

cules. Prog Nucl Magn Reson Spectrosc 2008;52(1):49-68.

methyl-TROSY view. J Magn Reson 2011;210(2):159-170.

some by NMR. Nature 2007;445(7128):618-622.

Prog Nucl Magn Reson Spectrosc 2010;56(3):247-266.

Molecular Biology 2012;831:133-140.

2009;38(2):129-143.

2009;44(4):213-223.

NMR. Top Curr Chem 2012;326:47-67.

2010;5(3):574-587.

403-410.


[102] Wang H, Peng D, Xie J. Ginseng leaf-stem: bioactive constituents and pharmacologi‐ cal functions. Chin Med 2009;4:20.

[88] Dalvit C, Fogliatto G, Stewart A, Veronesi M, Stockman B. WaterLOGSY as a method for primary NMR screening: practical aspects and range of applicability. J Biomol

[89] Balaram P, Bothner-By A, Breslow E. Localization of tyrosine at the binding site of neurophysin II by negative nuclear Overhouser effects. J Am Chem Soc 1972;94(11):

[90] Hajduk PJ, Olejniczak ET, Fesik SW. One-Dimensional Relaxation- and Diffusion-Edited NMR Methods for Screening Compounds That Bind to Macromolecules. J Am

[91] Kramer M, Kleinpeter E. STD-DOSY: A new NMR method to analyze multi-compo‐

[92] Fejzo J, Lepre C, Xie X. Application of NMR screening in drug discovery. Curr Top

[93] Lepre CA. Strategies for NMR screening and library design. Methods Princ Med

[94] Carlomagno T. Ligand-target interactions: what can we learn from NMR? Annu Rev

[95] Klages J, Coles M, Kessler H. NMR-based screening: a powerful tool in fragment-

[96] Murali N, Jarori GK, Landy SB, Rao BD. Two-dimensional transferred nuclear Over‐ hauser effect spectroscopy (TRNOESY) studies of nucleotide conformations in crea‐ tine kinase complexes: effects due to weak nonspecific binding. Biochemistry

[97] Koskela H, Heikkinen O, Kilpelainen I, Heikkinen S. Rapid and accurate processing method for amide proton exchange rate measurement in proteins. J Biomol NMR

[98] Howard GC, Brown WE, Editors. Modern Protein Chemistry: Practical Aspects. :

[99] Pauli GF, Jaki B, Lankin D, Burton I, Walter J. Quantitative NMR (qNMR) of Bioac‐ tive Natural Products. In: Colegate S, Molyneux R, editors. Bioactive Natural Prod‐ ucts: Detection, Isolation and Structural Determination. 2nd ed. Boca Raton, Florida:

[100] Merkley N, Syvitski RT. Profiling whole microalgal cells by high-resolution magic angle spinning (HR-MAS) magnetic resonance spectroscopy. J Appl Phycol

[101] Oliver SG. Functional genomics: lessons from yeast. Philos Trans R Soc Lond B Biol

nent enzyme/substrate systems. J Magn Reson 2010;202(2):245-249.

NMR 2001;21(4):349-359.

Chem 2003;16:391-415.

1993;32(47):12941-12948.

CRC Press; scheduled 2007.

2007;37(4):313-320.

2012;24(3):535-540.

Sci 2002;357(1417):17-23.

CRC; 2002.

Chem Soc 1997;119(50):12257-12261.

Med Chem (Hilversum, Neth ) 2003;3(1):81-97.

88 Using Old Solutions to New Problems - Natural Drug Discovery in the 21st Century

based drug discovery. Analyst 2007;132(7):693-705.

Biophys Biomol Struct 2005;34:245-266.

4017-4018.


receptor, reduces uptake of the contrast agent GdDOTA by murine orthotopic B16/BL6 melanoma tumours and inhibits their growth in vivo. NMR Biomed 2005;18(5):308-321.

[130] Rudin M, Allegrini P, Beckmann N, Gremlich HU, Kneuer R, Laurent D, et al. Nonin‐ vasive imaging in drug discovery and development. Ernst Schering Res Found Workshop 2004;(48)(48):47-75.

[117] Manach C, Hubert J, Llorach R, Scalbert A. The complex links between dietary phy‐ tochemicals and human health deciphered by metabolomics. Mol Nutr Food Res

[118] Power WP. High-resolution magic angle spinning-enabling applications of NMR spectroscopy to semi-solid phases. Annu Rep NMR Spectrosc 2011;72:111-156. [119] Weybright P, Millis K, Campbell N, Cory DG, Singer S. Gradient, high-resolution, magic angle spinning 1H nuclear magnetic resonance spectroscopy of intact cells.

[120] Lindon JC, Beckonert OP, Holmes E, Nicholson JK. High-resolution magic angle spinning NMR spectroscopy: Application to biomedical studies. Prog Nucl Magn Re‐

[121] Beckonert O, Coen M, Keun HC, Wang Y, Ebbels TM, Holmes E, et al. High-resolu‐ tion magic-angle-spinning NMR spectroscopy for metabolic profiling of intact tis‐

[122] Sitter B, Bathen TF, Tessem M, Gribbestad IS. High-resolution magic angle spinning (HR MAS) MR spectroscopy in metabolic characterization of human cancer. Prog

[123] Hanoulle X, Wieruszeski JM, Rousselot-Pailley P, Landrieu I, Baulard AR, Lippens G. Monitoring of the ethionamide pro-drug activation in mycobacteria by (1)H high res‐ olution magic angle spinning NMR. Biochem Biophys Res Commun 2005;331(2):

[124] Schroder H. High resolution magic angle spinning NMR for analyzing small mole‐ cules attached to solid support. Comb Chem High Throughput Screen 2003;6(7):

[125] Li W. Multidimensional HRMAS NMR: a platform for in vivo studies using intact

[126] Thongsaard W, Marsden C, Morris P, Prior M, Shah Y. Effect of Thunbergia laurifo‐ lia , a Thai natural product used to treat drug addiction, on cerebral activity detected by functional magnetic resonance imaging in the rat. Psychopharmacology (Berl )

[127] Miskowiak KW, Favaron E, Hafizi S, Inkster B, Goodwin GM, Cowen PJ, et al. Eryth‐ ropoietin modulates neural and cognitive processing of emotional information in bi‐ omarker models of antidepressant drug action in depressed patients.

[128] Miskowiak KW, Favaron E, Hafizi S, Inkster B, Goodwin GM, Cowen PJ, et al. Effects of erythropoietin on emotional processing biases in patients with major depression:

an exploratory fMRI study. Psychopharmacology (Berl) 2009;207(1):133-142. [129] Rudin M, McSheehy PM, Allegrini PR, Rausch M, Baumann D, Becquet M, et al. PTK787/ZK222584, a tyrosine kinase inhibitor of vascular endothelial growth factor

2009;53(10):1303-1315.

Magn Reson Med 1998;39(3):337-345.

90 Using Old Solutions to New Problems - Natural Drug Discovery in the 21st Century

sues. Nat Protoc 2010;5(6):1019-1032.

Nucl Magn Reson Spectrosc 2009;54(3-4):239-254.

bacterial cells. Analyst 2006;131(7):777-781.

Psychopharmacology (Berl) 2010;210(3):419-428.

son Spectrosc 2009;55(2):79-100.

452-458.

741-753.

2005;180(4):1-9.


**Bioactive Proteins from Natural Sources**
