Precursor-Based Selective Methyl Labeling of Cell-Free Synthesized Proteins

2018 ◽  
Vol 13 (8) ◽  
pp. 2170-2178 ◽  
Author(s):  
Mariya Lazarova ◽  
Frank Löhr ◽  
Ralf-Bernhardt Rues ◽  
Robin Kleebach ◽  
Volker Dötsch ◽  
...  
Keyword(s):  
Methyl Labeling

scholarly journals Improved strategy for isoleucine 1H/13C methyl labeling in Pichia pastoris

2019 ◽  
Vol 73 (12) ◽  
pp. 687-697 ◽  
Author(s):  
Rustam Ali ◽  
Lindsay D. Clark ◽  
Jacob A. Zahm ◽  
Andrew Lemoff ◽  
Karthik Ramesh ◽  
...  
Keyword(s):  
Pichia Pastoris ◽  
Isotope Labeling ◽  
Protein Complexes ◽  
Cost Effective ◽  
Specific Protein ◽  
Site Specific ◽  
Methyl Labeling ◽  
Structure And Dynamics ◽  
Eukaryotic Proteins ◽  
The Cost

Abstract Site specific methyl labeling combined with methyl TROSY offers a powerful NMR approach to study structure and dynamics of proteins and protein complexes of high molecular weight. Robust and cost-effective methods have been developed for site specific protein 1H/13C methyl labeling in an otherwise deuterated background in bacteria. However, bacterial systems are not suitable for expression and isotope labeling of many eukaryotic and membrane proteins. The yeast Pichia pastoris (P. pastoris) is a commonly used host for expression of eukaryotic proteins, and site-specific methyl labeling of perdeuterated eukaryotic proteins has recently been achieved with this system. However, the practical utility of methyl labeling and deuteration in P. pastoris is limited by high costs. Here, we describe an improved method for 1H/13C-labeling of the δ-methyl group of isoleucine residues in a perdeuterated background, which reduces the cost by ≥ 50% without compromising the efficiency of isotope enrichment. We have successfully implemented this method to label actin and a G-protein coupled receptor. Our approach will facilitate studies of the structure and dynamics of eukaryotic proteins by NMR spectroscopy.

scholarly journals Selective Methyl Labeling of Eukaryotic Membrane Proteins Using Cell-Free Expression

2014 ◽  
Vol 136 (32) ◽  
pp. 11308-11310 ◽  
Author(s):  
Rasmus Linser ◽  
Vladimir Gelev ◽  
Franz Hagn ◽  
Haribabu Arthanari ◽  
Sven G. Hyberts ◽  
...  
Keyword(s):  
Membrane Proteins ◽  
Free Expression ◽  
Methyl Labeling ◽  
Eukaryotic Membrane Proteins

scholarly journals Protein−Protein Interfaces Probed by Methyl Labeling and Proton-Detected Solid-State NMR Spectroscopy

ChemPhysChem ◽  
2018 ◽  
Vol 19 (19) ◽  
pp. 2457-2460 ◽  
Author(s):  
Maximilian Zinke ◽  
Pascal Fricke ◽  
Sascha Lange ◽  
Sophie Zinn-Justin ◽  
Adam Lange
Keyword(s):  
Solid State ◽  
Nmr Spectroscopy ◽  
Solid State Nmr ◽  
Solid State Nmr Spectroscopy ◽  
Methyl Labeling ◽  
Protein Interfaces

scholarly journals Erratum to: Methyl labeling and TROSY NMR spectroscopy of proteins expressed in the eukaryote Pichia pastoris

2016 ◽  
Vol 64 (3) ◽  
pp. 267-267
Author(s):  
Lindsay Clark ◽  
Jacob A. Zahm ◽  
Rustam Ali ◽  
Maciej Kukula ◽  
Liangqiao Bian ◽  
...  
Keyword(s):  
Nmr Spectroscopy ◽  
Pichia Pastoris ◽  
Methyl Labeling

Methyl labeling of HeLa cell hnRNA: a comparison with mRNA

Cell ◽  
1976 ◽  
Vol 7 (2) ◽  
pp. 227-237 ◽  
Author(s):  
M. Salditt-Georgieff ◽  
Warren Jelinek ◽  
James E. Darnell ◽  
Y. Furuichi ◽  
M. Morgan ◽  
...  
Keyword(s):  
Hela Cell ◽  
Methyl Labeling

Comparison of 13CH3, 13CH2D, and 13CHD2 methyl labeling strategies in proteins

2005 ◽  
Vol 33 (1) ◽  
pp. 25-41 ◽  
Author(s):  
Jason E. Ollerenshaw ◽  
Vitali Tugarinov ◽  
Nikolai R. Skrynnikov ◽  
Lewis E. Kay
Keyword(s):  
Methyl Labeling ◽  
Labeling Strategies

scholarly journals Methyl-Labeling Assisted NMR Structure Determination of a 66 KDA Growth Factor-Receptor Complex

2017 ◽  
Vol 112 (3) ◽  
pp. 487a-488a
Author(s):  
Andrew Hinck ◽  
Morkos A. Henen ◽  
Christian Zwieb ◽  
Ravindra Kodali ◽  
Cynthia S. Hinck
Keyword(s):  
Growth Factor ◽  
Structure Determination ◽  
Growth Factor Receptor ◽  
Nmr Structure ◽  
Receptor Complex ◽  
Nmr Structure Determination ◽  
Methyl Labeling
Sign in / Sign up

Export Citation Format

Share Document