Dictyostelium discoideum as Expression Host: Isotopic Labeling of a Recombinant Glycoprotein for NMR Studies

2000 ◽  
Vol 19 (3) ◽  
pp. 335-342 ◽  
Author(s):  
Liza Cubeddu ◽  
Catherine X. Moss ◽  
James D. Swarbrick ◽  
Andrew A. Gooley ◽  
Keith L. Williams ◽  
...  
Keyword(s):  
Dictyostelium Discoideum ◽  
Isotopic Labeling ◽  
Nmr Studies ◽  
Recombinant Glycoprotein ◽  
Expression Host

Recombinant glycoprotein production in the slime mould Dictyostelium discoideum

1995 ◽  
Vol 6 (5) ◽  
pp. 538-542 ◽  
Author(s):  
Keith L. Williams ◽  
Kerry R. Emslie ◽  
Martin B Slade
Keyword(s):  
Dictyostelium Discoideum ◽  
Recombinant Glycoprotein ◽  
Slime Mould

scholarly journals Robust Cell-Free Expression of Sub-Pathological and Pathological Huntingtin Exon-1 for NMR Studies. General Approaches for the Isotopic Labeling of Low-Complexity Proteins

Biomolecules ◽  
2020 ◽  
Vol 10 (10) ◽  
pp. 1458
Author(s):  
Anna Morató ◽  
Carlos A. Elena-Real ◽  
Matija Popovic ◽  
Aurélie Fournet ◽  
Karen Zhang ◽  
...  
Keyword(s):  
Protein Expression ◽  
Expression System ◽  
Isotopic Labeling ◽  
Low Complexity ◽  
Nonsense Suppression ◽  
Sequential Assignment ◽  
Reaction Yield ◽  
Nmr Studies ◽  
Intrinsically Disordered ◽  
Exon 1

The high-resolution structural study of huntingtin exon-1 (HttEx1) has long been hampered by its intrinsic properties. In addition to being prone to aggregate, HttEx1 contains low-complexity regions (LCRs) and is intrinsically disordered, ruling out several standard structural biology approaches. Here, we use a cell-free (CF) protein expression system to robustly and rapidly synthesize (sub-) pathological HttEx1. The open nature of the CF reaction allows the application of different isotopic labeling schemes, making HttEx1 amenable for nuclear magnetic resonance studies. While uniform and selective labeling facilitate the sequential assignment of HttEx1, combining CF expression with nonsense suppression allows the site-specific incorporation of a single labeled residue, making possible the detailed investigation of the LCRs. To optimize CF suppression yields, we analyze the expression and suppression kinetics, revealing that high concentrations of loaded suppressor tRNA have a negative impact on the final reaction yield. The optimized CF protein expression and suppression system is very versatile and well suited to produce challenging proteins with LCRs in order to enable the characterization of their structure and dynamics.

Alternate-site isotopic labeling of ribonucleotides for NMR studies of ribose conformational dynamics in RNA

2006 ◽  
Vol 35 (4) ◽  
pp. 261-274 ◽  
Author(s):  
James E. Johnson ◽  
Kristine R. Julien ◽  
Charles G. Hoogstraten
Keyword(s):  
Conformational Dynamics ◽  
Isotopic Labeling ◽  
Nmr Studies

Site-specific isotopic labeling of proteins for NMR studies

1992 ◽  
Vol 114 (20) ◽  
pp. 7959-7961 ◽  
Author(s):  
Jonathan A. Ellman ◽  
Brian F. Volkman ◽  
David Mendel ◽  
Peter G. Schulz ◽  
David E. Wemmer
Keyword(s):  
Isotopic Labeling ◽  
Nmr Studies ◽  
Site Specific

Dictyostelium discoideum as an expression host for the circumsporozoite protein of Plasmodium falciparum

Gene ◽  
1992 ◽  
Vol 111 (2) ◽  
pp. 157-163 ◽  
Author(s):  
Nicolas Fasel ◽  
Carole Begdadi-Rais ◽  
Michel Bernard ◽  
Claude Bron ◽  
Giampietro Corradin ◽  
...  
Keyword(s):  
Plasmodium Falciparum ◽  
Dictyostelium Discoideum ◽  
Circumsporozoite Protein ◽  
Expression Host

scholarly journals On the use of Pichia pastoris for isotopic labeling of human GPCRs for NMR studies

2018 ◽  
Vol 71 (4) ◽  
pp. 203-211 ◽  
Author(s):  
Lindsay Clark ◽  
Igor Dikiy ◽  
Daniel M. Rosenbaum ◽  
Kevin H. Gardner
Keyword(s):  
Pichia Pastoris ◽  
Isotopic Labeling ◽  
Nmr Studies

scholarly journals Chain-Selective Isotopic Labeling for NMR Studies of Large Multimeric Proteins: Application to Hemoglobin

2000 ◽  
Vol 79 (2) ◽  
pp. 1146-1154 ◽  
Author(s):  
Virgil Simplaceanu ◽  
Jonathan A. Lukin ◽  
Tsuei-Yun Fang ◽  
Ming Zou ◽  
Nancy T. Ho ◽  
...  
Keyword(s):  
Isotopic Labeling ◽  
Nmr Studies

Expression, purification and characterisation of secreted recombinant glycoprotein PsA in Dictyostelium discoideum

1995 ◽  
Vol 38 (2) ◽  
pp. 137-149 ◽  
Author(s):  
Ti Zhou-Chou ◽  
Martin B. Slade ◽  
Keith L. Williams ◽  
Andrew A. Gooley
Keyword(s):  
Dictyostelium Discoideum ◽  
Recombinant Glycoprotein

scholarly journals Methyl-specific isotopic labeling: a molecular tool box for solution NMR studies of large proteins

2015 ◽  
Vol 32 ◽  
pp. 113-122 ◽  
Author(s):  
Rime Kerfah ◽  
Michael J Plevin ◽  
Remy Sounier ◽  
Pierre Gans ◽  
Jerome Boisbouvier
Keyword(s):  
Isotopic Labeling ◽  
Solution Nmr ◽  
Nmr Studies ◽  
Large Proteins ◽  
Molecular Tool
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