Related Articles13C-NMR investigation of protein synthesis during apoptosis in human leukemic cell lines.
J Cell Physiol. 1999 Oct;181(1):147-52
Authors: Scott CE, Adebodun F
In order to evaluate the role of protein synthesis in apoptosis, (13)C-NMR has been used to study the levels of protein synthesis in three different human leukemic cell lines in the presence and absence of dexamethasone-induced apoptosis. Measurements were done on one dexamethasone-sensitive (CEM-C7-14) and two different dexamethasone-resistant variants (CEM-4R4 and CEM-ICR27-4). The incorporation of (13)C-labeled amino acids into cellular proteins, which reflects the level of new protein synthesis, was monitored by (13)C-NMR spectroscopy. In the absence of dexamethasone, the level of protein synthesis was found to be significantly different among the three cell lines. Dexamethasone caused a significant reduction ( congruent with 60-87%) in the level of protein synthesis in dexamethasone-sensitive CEM-C7-14 cells, while having no significant effect on protein synthesis in dexamethasone-resistant CEM-4R4 cells. Dexamethasone treatment caused a significant enhancement of the level of protein synthesis in the CEM-ICR27-4 cells. Synthesis of proteins was found to occur during apoptosis, albeit at a low level, suggesting a role for the synthesis of specific proteins in the mechanism of apoptosis.
Suppression of isotope scrambling in cell-free protein synthesis by broadband inhibition of PLP enymes for selective 15N-labelling and production of perdeuterated proteins in H2O
Suppression of isotope scrambling in cell-free protein synthesis by broadband inhibition of PLP enymes for selective 15N-labelling and production of perdeuterated proteins in H2O
Abstract Selectively isotope labelled protein samples can be prepared in vivo or in vitro from selectively labelled amino acids but, in many cases, metabolic conversions between different amino acids result in isotope scrambling. The best results are obtained by cell-free protein synthesis, where metabolic enzymes are generally less active, but isotope scrambling can never be suppressed completely. We show that...
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[NMR paper] Cell-free synthesis of 15N-labeled proteins for NMR studies.
Cell-free synthesis of 15N-labeled proteins for NMR studies.
Related Articles Cell-free synthesis of 15N-labeled proteins for NMR studies.
IUBMB Life. 2005 Sep;57(9):615-22
Authors: Ozawa K, Dixon NE, Otting G
Modern cell-free in vitro protein synthesis systems present powerful tools for the synthesis of isotope-labeled proteins in high yields. The production of selectively 15 N-labeled proteins from 15 N-labeled amino acids is particularly economic and yields are often sufficient to analyze the proteins very quickly by two-dimensional NMR...
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[NMR paper] Cell-free protein synthesis in an autoinduction system for NMR studies of protein-protein interactions.
Cell-free protein synthesis in an autoinduction system for NMR studies of protein-protein interactions.
Related Articles Cell-free protein synthesis in an autoinduction system for NMR studies of protein-protein interactions.
J Biomol NMR. 2005 Jul;32(3):235-41
Authors: Ozawa K, Jergic S, Crowther JA, Thompson PR, Wijffels G, Otting G, Dixon NA
Cell-free protein synthesis systems provide facile access to proteins in a nascent state that enables formation of soluble, native protein-protein complexes even if one of the protein components is prone...
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[NMR paper] Optimization of an Escherichia coli system for cell-free synthesis of selectively N-l
Optimization of an Escherichia coli system for cell-free synthesis of selectively N-labelled proteins for rapid analysis by NMR spectroscopy.
Related Articles Optimization of an Escherichia coli system for cell-free synthesis of selectively N-labelled proteins for rapid analysis by NMR spectroscopy.
Eur J Biochem. 2004 Oct;271(20):4084-93
Authors: Ozawa K, Headlam MJ, Schaeffer PM, Henderson BR, Dixon NE, Otting G
Cell-free protein synthesis offers rapid access to proteins that are selectively labelled with amino acids and suitable for...
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[NMR paper] NMR structure and mutagenesis of the inhibitor-of-apoptosis protein XIAP.
NMR structure and mutagenesis of the inhibitor-of-apoptosis protein XIAP.
Related Articles NMR structure and mutagenesis of the inhibitor-of-apoptosis protein XIAP.
Nature. 1999 Oct 21;401(6755):818-22
Authors: Sun C, Cai M, Gunasekera AH, Meadows RP, Wang H, Chen J, Zhang H, Wu W, Xu N, Ng SC, Fesik SW
The inhibitor-of-apoptosis (IAP) family of proteins, originally identified in baculoviruses, regulate programmed cell death in a variety of organisms. IAPs inhibit specific enzymes (caspases) in the death cascade and contain one to three...
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[NMR paper] Cell-free synthesis and amino acid-selective stable isotope labeling of proteins for
Cell-free synthesis and amino acid-selective stable isotope labeling of proteins for NMR analysis.
Related Articles Cell-free synthesis and amino acid-selective stable isotope labeling of proteins for NMR analysis.
J Biomol NMR. 1995 Sep;6(2):129-34
Authors: Kigawa T, Muto Y, Yokoyama S
For the application of multidimensional NMR spectroscopy to larger proteins, it would be useful to perform selective labeling of one of the 20 amino acids. For some amino acids, however, amino acid metabolism drastically reduces the efficiency and selectivity...
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Cell-free protein synthesis of perdeuterated proteins for NMR studies
Cell-free protein synthesis of perdeuterated proteins for NMR studies
Touraj Etezady-Esfarjani, Sebastian Hiller, Cristina Villalba and Kurt Wüthrich
Journal of Biomolecular NMR; 2007; 39(3); pp 229-238
Abstract:
Cell-free protein synthesis protocols for uniformly deuterated proteins typically yield low, non-uniform deuteration levels. This paper introduces an E. coli cell-extract, D-S30, which enables efficient production of proteins with high deuteration levels for all non-labile hydrogen atom positions. Potential applications of the new protocol may include production of proteins...