| Site-selective screening by NMR spectroscopy with labeled amino acid pairs. | |
| | |
MedLine Citation:
|
PMID: 11890787 Owner: NLM Status: MEDLINE |
Abstract/OtherAbstract:
|
A new method for site-selective screening by NMR is presented. The core of the new method is the dual amino acid sequence specific labeling technique. Amino acid X is labeled with (13)C and amino acid Y is labeled with (15)N. Provided only one XY pair occurs in the amino acid sequence, only one signal in the 1D carbonyl (13)C spectrum will display a splitting due to the (1)J(C'N) coupling. Using this labeling strategy it is possible to screen selectively for binding to a selected epitope without the need for sequence specific assignments. An HNCO spectrum (1D or 2D) can be used either directly as a screening experiment or indirectly to identify what signals to monitor in a 2D (1)H-(15)N correlation spectrum. Chemical shift perturbations upon addition of a potential ligand are easily detected even for large proteins due to the reduced spectral complexity resulting from the use of a selectively labeled sample. The new technique is demonstrated on the human adipocyte fatty acid binding protein FABP-4. Due to the reduced spectral complexity, the method should be applicable to larger proteins than are conventional methods. |
| | |
Authors:
|
Johan Weigelt; Maria van Dongen; Jonas Uppenberg; Johan Schultz; Mats Wikström |
Related Documents
:
|
10701257 - Development of an amino acid sequence and d/l-configuration determination method of pep... 11288727 - Amino acid sequence of a neurotoxic phospholipase a2 enzyme from common death adder (ac... 1662607 - Purification of two forms of arachidonate 15-lipoxygenase from human leukocytes. 10788797 - Activation of phospholipase a(2) by long chain fatty acyl groups involves a novel unsta... 9291117 - Biosynthesis of the labdane diterpene marrubiin in marrubium vulgare via a non-mevalona... 15156047 - Meat-model system development for antibacterial activity determination. |
Publication Detail:
|
Type: Journal Article |
Journal Detail:
|
Title: Journal of the American Chemical Society Volume: 124 ISSN: 0002-7863 ISO Abbreviation: J. Am. Chem. Soc. Publication Date: 2002 Mar |
Date Detail:
|
Created Date: 2002-03-13 Completed Date: 2002-05-23 Revised Date: - |
Medline Journal Info:
|
Nlm Unique ID: 7503056 Medline TA: J Am Chem Soc Country: United States |
Other Details:
|
Languages: eng Pagination: 2446-7 Citation Subset: IM |
Affiliation:
|
Department of Structural Chemistry, Biovitrum, S-112 76 Stockholm, Sweden. |
Export Citation:
|
APA/MLA Format Download EndNote Download BibTex |
| MeSH Terms | |
Descriptor/Qualifier:
|
Amino Acids
/
analysis* Binding Sites Carbon Isotopes Nitrogen Isotopes Nuclear Magnetic Resonance, Biomolecular / methods* Substrate Specificity |
| Chemical | |
Reg. No./Substance:
|
0/Amino Acids; 0/Carbon Isotopes; 0/Nitrogen Isotopes |
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine
Previous Document: Sensitive and rapid analysis of protein palmitoylation with a synthetic cell-permeable mimic of SRC ...
Next Document: Quantum dot on a rope.