Related ArticlesExpression and solution NMR study of multi-site phosphomimetic mutant BCL-2 protein.
Protein Pept Lett. 2019 Mar 27;:
Authors: Song T, Cao K, Fan Y, Zhang Z, Guo Z, Zhang M, Liu P
Abstract
The significance of multi-site phosphorylation of BCL-2 protein in the flexible loop domain remains controversial, in part due to the lack of structural biology studies of phosphorylated BCL-2. We constructed a phosphomietic mutant BCL-2(62-206) (t69e, s70e and s87e) (EEE-BCL-2-EK(62-206)), in which the BH4 domain and the part of loop region was trunkated (residues 2-61). In this way,we obtained a mass of soluble and stable phosphomimetic proteins to produce a well dispersed 15N-1H heteronuclear single quantum coherence (HSQC) NMR spectroscopy. The mutant protein reproduced the biochemical and cellular activity of the native phosphorylated BCL-2 (pBCL-2), which was distinct from nonphosphorylated BCL-2 (npBCL-2) protein. The difference between BH3 groove of EEE-BCL-2-EK(62-206) and BCL-2 protein in two-dimensional NMR spectra demonstrated that the phosphorylation induced structural change in active domain.This explains that phosphorylation regulates the anti-apoptotic function of BCL-2.
PMID: 30919764 [PubMed - as supplied by publisher]
[NMR paper] Protein-excipient interactions evaluated via NMR studies in polysorbate based multi-dose protein formulations: influence on antimicrobial efficacy and potential study approach.
Protein-excipient interactions evaluated via NMR studies in polysorbate based multi-dose protein formulations: influence on antimicrobial efficacy and potential study approach.
Related Articles Protein-excipient interactions evaluated via NMR studies in polysorbate based multi-dose protein formulations: influence on antimicrobial efficacy and potential study approach.
J Pharm Sci. 2018 Jun 05;:
Authors: Torosantucci R, Furtmann B, Elshorst B, Pfeiffer-Marek S, Hartleb T, Andres N, Bussemer T
Abstract
Preservatives are excipients...
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06-10-2018 03:09 AM
Protein-excipient interactions evaluated via NMR studies in polysorbate based multi-dose protein formulations: influence on antimicrobial efficacy and potential study approach
Protein-excipient interactions evaluated via NMR studies in polysorbate based multi-dose protein formulations: influence on antimicrobial efficacy and potential study approach
Publication date: Available online 5 June 2018
Source:Journal of Pharmaceutical Sciences</br>
Author(s): Riccardo Torosantucci, Britta Furtmann, Bettina Elshorst, Stefania Pfeiffer-Marek, Tanja Hartleb, Nikolaus Andres, Till Bussemer</br>
Preservatives are excipients essentially needed in pharmaceutical multi-dose formulations to prevent microbial growth. Among available...
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06-06-2018 01:40 AM
[NMR paper] Sequential Protein Expression and Capsid Assembly In Cell: Towards the Study of Multi-Protein Viral Capsids Using Solid-State NMR Techniques.
Sequential Protein Expression and Capsid Assembly In Cell: Towards the Study of Multi-Protein Viral Capsids Using Solid-State NMR Techniques.
Related Articles Sequential Protein Expression and Capsid Assembly In Cell: Towards the Study of Multi-Protein Viral Capsids Using Solid-State NMR Techniques.
Biochemistry. 2018 Feb 21;:
Authors: Alphonse S, Itin B, Khayat R, Ghose R
Abstract
While solid-state NMR (ssNMR) has emerged as a powerful technique to study viral capsids, current studies are limited to capsids formed from single...
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02-23-2018 03:44 AM
[NMR paper] Preparation of Multiple Site-Specific Mutant Proteins for NMR Studies by PCR-Directed Cell-Free Protein Synthesis.
Preparation of Multiple Site-Specific Mutant Proteins for NMR Studies by PCR-Directed Cell-Free Protein Synthesis.
Preparation of Multiple Site-Specific Mutant Proteins for NMR Studies by PCR-Directed Cell-Free Protein Synthesis.
Methods Mol Biol. 2014;1118:169-87
Authors: Ozawa K, Qi R
Abstract
Cell-free protein synthesis (CFPS) offers a fast and inexpensive approach to selectively label proteins with isotopes that can then be detected by nuclear magnetic resonance (NMR) spectroscopy directly in the translation mixture. We describe...
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01-08-2014 11:23 AM
Segmental isotope labeling of proteins for NMR structural study using a protein S tag for higher expression and solubility
Segmental isotope labeling of proteins for NMR structural study using a protein S tag for higher expression and solubility
Abstract A common obstacle to NMR studies of proteins is sample preparation. In many cases, proteins targeted for NMR studies are poorly expressed and/or expressed in insoluble forms. Here, we describe a novel approach to overcome these problems. In the protein S tag-intein (PSTI) technology, two tandem 92-residue N-terminal domains of protein S (PrS2) from Myxococcus xanthus is fused at the N-terminal end of a protein to enhance its expression and solubility. Using...
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03-08-2012 08:46 AM
[NMR paper] Solution 1H NMR study of the active site molecular structure and magnetic properties
Solution 1H NMR study of the active site molecular structure and magnetic properties of the cyanomet complex of the isolated, tetrameric beta-chain from human adult hemoglobin.
Related Articles Solution 1H NMR study of the active site molecular structure and magnetic properties of the cyanomet complex of the isolated, tetrameric beta-chain from human adult hemoglobin.
Biochim Biophys Acta. 2004 Sep 1;1701(1-2):75-87
Authors: Tran AT, Kolczak U, La Mar GN
The solution molecular structure and the electronic and magnetic properties of the heme...
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[NMR paper] Flavonoid binding to a multi-drug-resistance transporter protein: an STD-NMR study.
Flavonoid binding to a multi-drug-resistance transporter protein: an STD-NMR study.
Related Articles Flavonoid binding to a multi-drug-resistance transporter protein: an STD-NMR study.
Anal Bioanal Chem. 2004 Aug;379(7-8):1045-9
Authors: Nissler L, Gebhardt R, Berger S
Flavonoids are well known to inhibit the function of the multi-drug-resistance (mdr) transporter by interacting with their ATP binding domains. The precise orientation of these molecules inside the ATP binding pocket is still unclear. We applied the saturation transfer...
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[NMR paper] NMR study on solution structure of the site-specific mutant Leu48----Ala transforming
NMR study on solution structure of the site-specific mutant Leu48----Ala transforming growth factor alpha.
Related Articles NMR study on solution structure of the site-specific mutant Leu48----Ala transforming growth factor alpha.
Int J Pept Protein Res. 1992 Feb;39(2):111-6
Authors: Kline TP, Mueller L
The NMR spectra of the Leu48----Ala mutant of human transforming growth factor alpha were compared to that of the wild-type. All chemical shift changes are less than or equal to 0.02 ppm with the exception of resonances associated with residues...
Expression and solution NMR study of multi-site phosphomimetic mutant BCL-2 protein.
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