A collection of small molecules (MW < 350 Da) was screened for binding to human factor Xa using saturation transfer difference NMR spectroscopy to detect binding. The NMR screening experiments identified four hits. Binding isotherms constructed from NMR linewidth data showed that the binding affinities of the hits were all in the 30-210 microM range. Competition binding experiments showed that three of the ligands were displaced by a known microM inhibitor of factor Xa. The success of the method for identifying new ligands and the relevance of this information to the design of new factor Xa inhibitors are discussed.
Preparation of protein samples for NMR structure, function, and small-molecule screening studies.
Preparation of protein samples for NMR structure, function, and small-molecule screening studies.
Preparation of protein samples for NMR structure, function, and small-molecule screening studies.
Methods Enzymol. 2011;493:21-60
Authors: Acton TB, Xiao R, Anderson S, Aramini J, Buchwald WA, Ciccosanti C, Conover K, Everett J, Hamilton K, Huang YJ, Janjua H, Kornhaber G, Lau J, Lee DY, Liu G, Maglaqui M, Ma L, Mao L, Patel D, Rossi P, Sahdev S, Shastry R, Swapna GV, Tang Y, Tong S, Wang D, Wang H, Zhao L, Montelione GT
In this chapter, we...
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[NMR paper] Expression screening, protein purification and NMR analysis of human protein domains
Expression screening, protein purification and NMR analysis of human protein domains for structural genomics.
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J Struct Funct Genomics. 2004;5(1-2):119-31
Authors: Folkers GE, van Buuren BN, Kaptein R
Structural genomics, the determination of protein structures on a genome-wide scale, is still in its infancy for eukaryotes due to the number and size of their genes. Low protein expression and solubility of eukaryotic...
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[NMR paper] Second-site NMR screening and linker design.
Second-site NMR screening and linker design.
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Curr Top Med Chem. 2003;3(1):69-80
Authors: Jahnke W, Flörsheimer A, Blommers MJ, Paris CG, Heim J, Nalin CM, Perez LB
One of the prime merits of NMR as a tool for lead finding in drug discovery research is its sensitivity and robustness to detect weak protein-ligand interactions. This sensitivity allows to build up ligands for a given target in a modular way, by a fragment-based approach. In this approach, two ligands are seperately...
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[NMR paper] Exploring the calcium-binding site in photosystem II membranes by solid-state (113)Cd
Exploring the calcium-binding site in photosystem II membranes by solid-state (113)Cd NMR.
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Biochemistry. 2000 Jun 13;39(23):6751-5
Authors: Matysik J, Alia A, Nachtegaal G, van Gorkom HJ, Hoff AJ, de Groot HJ
Calcium (Ca(2+)) is an essential cofactor for photosynthetic oxygen evolution. Although the involvement of Ca(2+) at the oxidizing side of photosystem II of plants has been known for a long time, its ligand interactions and mode of...
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[NMR paper] Structural determination of the active site of a sweet protein. A 1H NMR investigatio
Structural determination of the active site of a sweet protein. A 1H NMR investigation of pMNEI.
http://www.ncbi.nlm.nih.gov/corehtml/query/egifs/http:--linkinghub.elsevier.com-ihub-images-PubMedLink.gif Related Articles Structural determination of the active site of a sweet protein. A 1H NMR investigation of pMNEI.
FEBS Lett. 1992 Sep 21;310(1):27-30
Authors: Tancredi T, Iijima H, Saviano G, Amodeo P, Temussi PA
pMNEI, a single chain sweet protein related to monellin, has been studied by means of 1H NMR at 500 MHz. A partial sequential...
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[NMR paper] Assignment of active-site protons in the 1H-NMR spectrum of reduced human Cu/Zn super
Assignment of active-site protons in the 1H-NMR spectrum of reduced human Cu/Zn superoxide dismutase.
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Eur J Biochem. 1991 May 8;197(3):691-7
Authors: Bertini I, Capozzi F, Luchinat C, Piccioli M, Viezzoli MS
600-MHz 1H-NMR and nuclear Overhauser enhancement spectroscopy (NOESY)...
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[NMR paper] Structure-function relationships in human epidermal growth factor studied by site-dir
Structure-function relationships in human epidermal growth factor studied by site-directed mutagenesis and 1H NMR.
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Biochemistry. 1991 Sep 10;30(36):8891-8
Authors: Hommel U, Dudgeon TJ, Fallon A, Edwards RM, Campbell ID
In order to elucidate the mechanism of interaction between human epidermal growth factor (EGF) and its receptor, selected variants of EGF, differing by single amino acid substitutions, have been...
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[NMR paper] Structure-function relationships in human epidermal growth factor studied by site-dir
Structure-function relationships in human epidermal growth factor studied by site-directed mutagenesis and 1H NMR.
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Biochemistry. 1991 Sep 10;30(36):8891-8
Authors: Hommel U, Dudgeon TJ, Fallon A, Edwards RM, Campbell ID
In order to elucidate the mechanism of interaction between human epidermal growth factor (EGF) and its receptor, selected variants of EGF, differing by single amino acid substitutions, have been...
Exploring the active site of human factor Xa protein by NMR screening of small molecu
Linear Mode
