scholarly journals Isotopic Labeling of Eukaryotic Membrane Proteins for NMR Studies of Interactions and Dynamics

2019 ◽  
pp. 37-65 ◽  
Author(s):  
Igor Dikiy ◽  
Lindsay D. Clark ◽  
Kevin H. Gardner ◽  
Daniel M. Rosenbaum
Keyword(s):  
Membrane Proteins ◽  
Isotopic Labeling ◽  
Nmr Studies ◽  
Eukaryotic Membrane Proteins

Synthetic Biology-Based Solution NMR Studies on Membrane Proteins in Lipid Environments

2019 ◽  
pp. 143-185 ◽  
Author(s):  
Erik Henrich ◽  
Frank Löhr ◽  
Julija Mezhyrova ◽  
Aisha Laguerre ◽  
Frank Bernhard ◽  
...  
Keyword(s):  
Synthetic Biology ◽  
Membrane Proteins ◽  
Solution Nmr ◽  
Nmr Studies

Lipid−Protein Nanodiscs as Reference Medium in Detergent Screening for High-Resolution NMR Studies of Integral Membrane Proteins

2010 ◽  
Vol 132 (16) ◽  
pp. 5628-5629 ◽  
Author(s):  
Zakhar O. Shenkarev ◽  
Ekaterina N. Lyukmanova ◽  
Alexander S. Paramonov ◽  
Lyudmila N. Shingarova ◽  
Vladimir V. Chupin ◽  
...  
Keyword(s):  
High Resolution ◽  
Membrane Proteins ◽  
Integral Membrane Proteins ◽  
Nmr Studies ◽  
High Resolution Nmr ◽  
Reference Medium

Regulation of the water channel aquaporin-2 by posttranslational modification

2011 ◽  
Vol 300 (5) ◽  
pp. F1062-F1073 ◽  
Author(s):  
Hanne B. Moeller ◽  
Emma T. B. Olesen ◽  
Robert A. Fenton
Keyword(s):  
Membrane Proteins ◽  
Protein Interactions ◽  
Posttranslational Modifications ◽  
Posttranslational Modification ◽  
Water Channel ◽  
Cellular Functions ◽  
Aquaporin 2 ◽  
Eukaryotic Membrane Proteins ◽  
Insight Into

The cellular functions of many eukaryotic membrane proteins, including the vasopressin-regulated water channel aquaporin-2 (AQP2), are regulated by posttranslational modifications. In this article, we discuss the experimental discoveries that have advanced our understanding of how posttranslational modifications affect AQP2 function, especially as they relate to the role of AQP2 in the kidney. We review the most recent data demonstrating that glycosylation and, in particular, phosphorylation and ubiquitination are mechanisms that regulate AQP2 activity, subcellular sorting and distribution, degradation, and protein interactions. From a clinical perspective, posttranslational modification resulting in protein misrouting or degradation may explain certain forms of nephrogenic diabetes insipidus. In addition to providing major insight into the function and dynamics of renal AQP2 regulation, the analysis of AQP2 posttranslational modification may provide general clues as to the role of posttranslational modification for regulation of other membrane proteins.

scholarly journals Understanding GPCR recognition and folding from NMR studies of fragments

RSC Advances ◽  
2018 ◽  
Vol 8 (18) ◽  
pp. 9858-9870 ◽  
Author(s):  
Jacopo Marino ◽  
Reto Walser ◽  
Martin Poms ◽  
Oliver Zerbe
Keyword(s):  
Protein Folding ◽  
Membrane Proteins ◽  
Membrane Insertion ◽  
Nmr Studies

Cotranslational protein folding is a vectorial process, and for membrane proteins, N-terminal helical segments are the first that become available for membrane insertion. Here fragments corresponding to these segments are investigated by NMR.

scholarly journals Codon Harmonization of a Kir3.1-KirBac1.3 Chimera for Structural Study Optimization

Biomolecules ◽  
2020 ◽  
Vol 10 (3) ◽  
pp. 430
Author(s):  
Evan Van Aalst ◽  
Maryam Yekefallah ◽  
Anil K. Mehta ◽  
Isaac Eason ◽  
Benjamin Wylie
Keyword(s):  
Membrane Proteins ◽  
G Protein ◽  
Recombinant Expression ◽  
Protein Product ◽  
Inward Rectifier ◽  
Correlation Spectroscopy ◽  
Protein Yield ◽  
K Channel ◽  
Functional Assay ◽  
Nmr Studies

The expression of functional, folded, and isotopically enriched membrane proteins is an enduring bottleneck for nuclear magnetic resonance (NMR) studies. Indeed, historically, protein yield optimization has been insufficient to allow NMR analysis of many complex Eukaryotic membrane proteins. However, recent work has found that manipulation of plasmid codons improves the odds of successful NMR-friendly protein production. In the last decade, numerous studies showed that matching codon usage patterns in recombinant gene sequences to those in the native sequence is positively correlated with increased protein yield. This phenomenon, dubbed codon harmonization, may be a powerful tool in optimizing recombinant expression of difficult-to-produce membrane proteins for structural studies. Here, we apply this technique to an inward rectifier K+ Channel (Kir) 3.1-KirBac1.3 chimera. Kir3.1 falls within the G protein-coupled inward rectifier K+ (GIRK) channel family, thus NMR studies may inform on the nuances of GIRK gating action in the presence and absence of its G Protein, lipid, and small molecule ligands. In our hands, harmonized plasmids increase protein yield nearly two-fold compared to the traditional ‘fully codon optimized’ construct. We then employ a fluorescence-based functional assay and solid-state NMR correlation spectroscopy to show the final protein product is folded and functional.

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