Abstract
Protein-Observed Fluorine NMR Spectroscopy (PrOF NMR) is an emerging technique for screening and characterizing small molecule-protein interactions. The choice of which amino acid to label for PrOF NMR can be critical for analysis. Here we report the first use of a protein containing two different fluoroaromatic amino acids for NMR studies. Using the KIX domain of the CBP/p300 as a model system, we examine ligand binding of several small molecules elaborated from our previous fragment screen and identify a new ligand binding site distinct from those used by native transcription factors. This site was further supported by computational modeling (FTMap and Schrödinger) and 1H-15N HSQC/HMQC NMR spectroscopy. Metabolic labelling with multiple fluorinated amino acids provides useful probes for further studying ligand binding and has led to new insight for allosterically regulating transcription-factor protein interactions with small molecules.
PMID: 29430847 [PubMed - as supplied by publisher]
[NMR paper] Acceleration of protein backbone NMR assignment by combinatorial labeling: Application to a small molecule binding study.
Acceleration of protein backbone NMR assignment by combinatorial labeling: Application to a small molecule binding study.
Acceleration of protein backbone NMR assignment by combinatorial labeling: Application to a small molecule binding study.
Biopolymers. 2016 Dec 30;:
Authors: Hein C, Löhr F, Schwarz D, Dötsch V
Abstract
Selective labeling with stable isotopes has long been recognized as a valuable tool in protein NMR to alleviate signal overlap and sensitivity limitations. In this study, combinatorial (15) N-, (13) C(?) -,...
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[NMR paper] Identification of Small-Molecule Inhibitors of the HuR/RNA Interaction Using a Fluorescence Polarization Screening Assay Followed by NMR Validation.
Identification of Small-Molecule Inhibitors of the HuR/RNA Interaction Using a Fluorescence Polarization Screening Assay Followed by NMR Validation.
Related Articles Identification of Small-Molecule Inhibitors of the HuR/RNA Interaction Using a Fluorescence Polarization Screening Assay Followed by NMR Validation.
PLoS One. 2015;10(9):e0138780
Authors: Wang Z, Bhattacharya A, Ivanov DN
Abstract
The human antigen R (HuR) stabilizes many mRNAs of proto-oncogene, transcription factors, cytokines and growth factors by recognizing...
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09-22-2015 06:40 PM
[NMR paper] TROSY NMR with a 52 kDa sugar transport protein and the binding of a small-molecule inhibitor.
TROSY NMR with a 52 kDa sugar transport protein and the binding of a small-molecule inhibitor.
Related Articles TROSY NMR with a 52 kDa sugar transport protein and the binding of a small-molecule inhibitor.
Mol Membr Biol. 2014 May 7;
Authors: Kalverda AP, Gowdy J, Thompson GS, Homans SW, Henderson PJ, Patching SG
Abstract
Abstract Using the sugar transport protein, GalP, from Escherichia coli, which is a homologue of human GLUT transporters, we have overcome the challenges for achieving high-resolution - and -methyl-TROSY NMR...
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05-09-2014 07:01 PM
Small-Molecule Binding Sites on Proteins Establishedby Paramagnetic NMR Spectroscopy
Small-Molecule Binding Sites on Proteins Establishedby Paramagnetic NMR Spectroscopy
Jia-Ying Guan, Peter H. J. Keizers, Wei-Min Liu, Frank Lo?hr, Simon P. Skinner, Edwin A. Heeneman, Harald Schwalbe, Marcellus Ubbink and Gregg Siegal
http://pubs.acs.org/appl/literatum/publisher/achs/journals/content/jacsat/0/jacsat.ahead-of-print/ja401323m/aop/images/medium/ja-2013-01323m_0009.gif
Journal of the American Chemical Society
DOI: 10.1021/ja401323m
http://feeds.feedburner.com/~ff/acs/jacsat?d=yIl2AUoC8zA
http://feeds.feedburner.com/~r/acs/jacsat/~4/ZukfjIRmQq8
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[NMR paper] Small molecule binding sites on proteins established by paramagnetic NMR spectroscopy.
Small molecule binding sites on proteins established by paramagnetic NMR spectroscopy.
Related Articles Small molecule binding sites on proteins established by paramagnetic NMR spectroscopy.
J Am Chem Soc. 2013 Mar 20;
Authors: Guan JY, Keizers PH, Liu WM, Loehr F, Skinner SP, Heeneman EA, Schwalbe H, Ubbink M, Siegal GD
Abstract
Determining the three dimensional structure of a small molecule-protein complex with weak affinity can be a significant challenge. We present a paramagnetic NMR method to determine intermolecular structure...
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03-21-2013 02:58 PM
[NMR paper] NMR Methods for Detection of Small Molecule Binding to RGS4.
NMR Methods for Detection of Small Molecule Binding to RGS4.
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Methods Enzymol. 2013;522:133-52
Authors: Storaska AJ, Neubig RR
Abstract
The duration and amplitude of G-protein-coupled receptor (GPCR) signaling is controlled by regulator of G-protein signaling (RGS) proteins. The 20 RGS family members act as GTPase accelerating proteins through their interaction with the G? subunit of the G??? heterotrimer. Their influence over GPCR signaling has attracted many to...
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02-05-2013 09:51 PM
Two-dimensional concurrent HMQC-COSY as an approach for small molecule chemical shift assignment and compound identification
Two-dimensional concurrent HMQC-COSY as an approach for small molecule chemical shift assignment and compound identification
Abstract Chemical shift assignment is the first step toward the structure elucidation of natural products and other chemical compounds. We propose here the use of 2D concurrent HMQC-COSY as an experiment for rapid chemical shift assignment of small molecules. This experiment provides well-dispersed 1Hâ??13C peak patterns that are distinctive for different functional groups plus 1Hâ??1H COSY connectivities that serve to identify adjacent groups. The COSY diagonal...
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[NMR paper] Annexin V binding to the outer leaflet of small unilamellar vesicles leads to altered
Annexin V binding to the outer leaflet of small unilamellar vesicles leads to altered inner-leaflet properties: 31P- and 1H-NMR studies.
Related Articles Annexin V binding to the outer leaflet of small unilamellar vesicles leads to altered inner-leaflet properties: 31P- and 1H-NMR studies.
Biochemistry. 1994 Sep 13;33(36):10944-50
Authors: Swairjo MA, Roberts MF, Campos MB, Dedman JR, Seaton BA
Calcium-dependent binding to phospholipid membranes is closely associated with annexin functional properties. In these studies, 31P- and 1H-nuclear...
Dual Labeling of the CBP/p300 KIX domain for 19F NMR leads to identification of a new small molecule binding site.
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