12 research outputs found
Phosphorylation-induced conformation of beta(2)-adrenoceptor related to arrestin recruitment revealed by NMR
The C-terminal region of G-protein-coupled receptors (GPCRs), stimulated by agonist binding, is phosphorylated by GPCR kinases, and the phosphorylated GPCRs bind to arrestin, leading to the cellular responses. To understand the mechanism underlying the formation of the phosphorylated GPCR-arrestin complex, we performed NMR analyses of the phosphorylated beta(2)-adrenoceptor (beta(2)AR) and the phosphorylated beta(2)AR-beta-arrestin 1 complex, in the lipid bilayers of nanodisc. Here we show that the phosphorylated C-terminal region adheres to either the intracellular side of the transmembrane region or lipids, and that the phosphorylation of the C-terminal region allosterically alters the conformation around M215(5.54) and M279(6.41), located on transemembrane helices 5 and 6, respectively. In addition, we found that the conformation induced by the phosphorylation is similar to that corresponding to the beta-arrestin-bound state. The phosphorylation-induced structures revealed in this study propose a conserved structural motif of GPCRs that enables beta-arrestin to recognize dozens of GPCRs.Peer reviewe
Comprehensive Analytical Method for the Determination of Hydrophilic Metabolites by High-Performance Liquid Chromatography and Mass Spectrometry
Advantage of LC-MS Metabolomics Methodology Targeting Hydrophilic Compounds in the Studies of Fermented Food Samples
ATP-dependent chromatin remodeling facilitates nucleotide excision repair of UV-induced DNA lesions in synthetic dinucleosomes
To investigate the relationship between chromatin dynamics and nucleotide excision repair (NER), we have examined the effect of chromatin structure on the formation of two major classes of UV-induced DNA lesions in reconstituted dinucleosomes. Furthermore, we have developed a model chromatin-NER system consisting of purified human NER factors and dinucleosome substrates that contain pyrimidine (6-4) pyrimidone photoproducts (6-4PPs) either at the center of the nucleosome or in the linker DNA. We have found that the two classes of UV-induced DNA lesions are formed efficiently at every location on dinucleosomes in a manner similar to that of naked DNA, even in the presence of histone H1. On the other hand, excision of 6-4PPs is strongly inhibited by dinucleosome assembly, even within the linker DNA region. These results provide direct evidence that the human NER machinery requires a space greater than the size of the linker DNA to excise UV lesions efficiently. Interestingly, NER dual incision in dinucleosomes is facilitated by recombinant ACF, an ATP-dependent chromatin remodeling factor. Our results indicate that there is a functional connection between chromatin remodeling and the initiation step of NER
Crystallization and preliminary crystallographic analysis of the N-terminal domain of PriA from Escherichia coli
Structure and dynamic studies by NMR of the potent sweet protein monellin and a non-sweet analog
Interaction Analysis of FABP4 Inhibitors by X‑ray Crystallography and Fragment Molecular Orbital Analysis
X-ray crystal structural determination
of FABP4 in complex with
four inhibitors revealed the complex binding modes, and the resulting
observations led to improvement of the inhibitory potency of FABP4
inhibitors. However, the detailed structure–activity relationship
(SAR) could not be explained from these structural observations. For
a more detailed understanding of the interactions between FABP4 and
inhibitors, fragment molecular orbital analyses were performed. These
analyses revealed that the total interfragment interaction energies
of FABP4 and each inhibitor correlated with the ranking of the <i>K</i><sub>i</sub> value for the four inhibitors. Furthermore,
interactions between each inhibitor and amino acid residues in FABP4
were identified. The oxygen atom of Lys58 in FABP4 was found to be
very important for strong interactions with FABP4. These results might
provide useful information for the development of novel potent FABP4
inhibitors