Related ArticlesProtein NMR Studies of substrate binding to human blood group A and B glycosyltransferases.
Chembiochem. 2017 Mar 03;:
Authors: Peters T, Grimm LL, Weissbach S, Flügge F, Begemann N, Palcic M
Abstract
Donor and acceptor substrate binding to human blood group A and B glycosyltransferases (GTA, GTB) has been studied by a variety of protein NMR experiments. Prior crystallographic studies have shown these enzymes to adopt an open conformation in the absence of substrates. Binding of either the donor substrate UDP-Gal, or of UDP induces a semi-closed conformation. In the presence of both, donor- and acceptor substrates, the enzymes shift towards a closed conformation with ordering of an internal loop and the C-terminal residues, which then completely cover the donor-binding pocket. Chemical shift titrations of uniformly 2H,15N labeled GTA or GTB with UDP affected about 20% of all cross peaks in 1H,15N-TROSY-HSQC spectra reflecting substantial plasticity of the enzymes. On the other hand, it is this conformational flexibility that impedes NH backbone assignments. Chemical shift perturbation experiments using ?1-13C-methyl Ile labeled samples revealed two Ile residues, Ile123 at the bottom of the UDP binding pocket, and Ile192 as part of the internal loop that were significantly disturbed upon stepwise addition of UDP and H-disaccharide, also revealing long-range perturbations. Finally, methyl TROSY based relaxation dispersion experiments do not reveal ?s to ms time scale motions. Although this study reveals substantial conformational plasticity of GTA and GTB it remains enigmatic how binding of substrates shifts the enzymes into catalytically competent states.
PMID: 28256109 [PubMed - as supplied by publisher]
[NMR paper] Compact NMR relaxometry of human blood and blood components.
Compact NMR relaxometry of human blood and blood components.
Related Articles Compact NMR relaxometry of human blood and blood components.
Trends Analyt Chem. 2016 Nov;83(A):53-64
Authors: Cistola DP, Robinson MD
Abstract
Nuclear magnetic resonance relaxometry is a uniquely practical and versatile implementation of NMR technology. Because it does not depend on chemical shift resolution, it can be performed using low-field compact instruments deployed in atypical settings. Early relaxometry studies of human blood were focused on...
nmrlearner
Journal club
0
12-24-2016 08:34 AM
[NMR paper] A rigid lanthanide binding tag to aid NMR studies of a 70 kDa homodimeric coat protein of human norovirus.
A rigid lanthanide binding tag to aid NMR studies of a 70 kDa homodimeric coat protein of human norovirus.
Related Articles A rigid lanthanide binding tag to aid NMR studies of a 70 kDa homodimeric coat protein of human norovirus.
Chem Commun (Camb). 2015 Nov 10;
Authors: Mallagaray A, Domínguez G, Peters T, Pérez-Castells J
Abstract
Attachment of human noroviruses to histo blood group antigens is thought to be essential for infection of host cells. Molecular details of the attachment process can be studied in vitro using a...
[NMR paper] NMR structure of human restriction factor APOBEC3A reveals substrate binding and enzyme specificity.
NMR structure of human restriction factor APOBEC3A reveals substrate binding and enzyme specificity.
Related Articles NMR structure of human restriction factor APOBEC3A reveals substrate binding and enzyme specificity.
Nat Commun. 2013;4:1890
Authors: Byeon IJ, Ahn J, Mitra M, Byeon CH, Hercík K, Hritz J, Charlton LM, Levin JG, Gronenborn AM
Abstract
Human APOBEC3A is a single-stranded DNA cytidine deaminase that restricts viral pathogens and endogenous retrotransposons, and has a role in the innate immune response. Furthermore, its...
nmrlearner
Journal club
0
05-23-2013 06:54 PM
Bacterial expression, purification, and model membrane reconstitution of the transmembrane and cytoplasmic domains of the human APP binding protein LR11/SorLA for NMR studies.
Bacterial expression, purification, and model membrane reconstitution of the transmembrane and cytoplasmic domains of the human APP binding protein LR11/SorLA for NMR studies.
Bacterial expression, purification, and model membrane reconstitution of the transmembrane and cytoplasmic domains of the human APP binding protein LR11/SorLA for NMR studies.
Protein Expr Purif. 2011 Feb 11;
Authors: Wang X, Gill Jr RL, Zhu Q, Tian F
LR11 (SorLA) is a recently identified neuronal protein that interacts with amyloid precursor protein (APP), a central player...
[NMR paper] The substrate binding site of human liver cytochrome P450 2C9: an NMR study.
The substrate binding site of human liver cytochrome P450 2C9: an NMR study.
http://www.ncbi.nlm.nih.gov/corehtml/query/egifs/http:--pubs.acs.org-images-acspubs.jpg Related Articles The substrate binding site of human liver cytochrome P450 2C9: an NMR study.
Biochemistry. 1997 Oct 21;36(42):12672-82
Authors: Poli-Scaife S, Attias R, Dansette PM, Mansuy D
Purified recombinant human liver cytochrome P450 2C9 was produced, from expression of the corresponding cDNA in yeast, in quantities large enough for UV-visible and 1H NMR experiments. Its...
nmrlearner
Journal club
0
08-22-2010 05:08 PM
[NMR paper] 1H NMR studies of reactions of copper complexes with human blood plasma and urine.
1H NMR studies of reactions of copper complexes with human blood plasma and urine.
Related Articles 1H NMR studies of reactions of copper complexes with human blood plasma and urine.
Biochem Pharmacol. 1992 Jan 22;43(2):137-45
Authors: Bligh SW, Boyle HA, McEwen AB, Sadler PJ, Woodham RH
Reactions of the copper complexes Cu(II)Cl2, 2-, and + (where DIPS is 3,5-diisopropylsalicylate and DMP is 2,9-dimethylphenanthroline) with human blood plasma and urine have been studied by 500 MHz 1H NMR spectroscopy, and CD spectroscopy has been used to...
Protein NMR Studies of substrate binding to human blood group A and B glycosyltransferases.
Linear Mode
