Faculty Opinions recommendation of Segmental isotopic labeling of a central domain in a multidomain protein by protein trans-splicing using only one robust DnaE intein.

2009 ◽  
Author(s):  
David Cowburn
Keyword(s):  
Isotopic Labeling ◽  
Central Domain ◽  
Multidomain Protein ◽  
Trans Splicing ◽  
Segmental Isotopic Labeling

Segmental Isotopic Labeling of a Central Domain in a Multidomain Protein by ProteinTrans-Splicing Using Only One Robust DnaE Intein

2009 ◽  
Vol 48 (33) ◽  
pp. 6128-6131 ◽  
Author(s):  
Alena E. L. Busche ◽  
A. Sesilja Aranko ◽  
Mehdi Talebzadeh-Farooji ◽  
Frank Bernhard ◽  
Volker Dötsch ◽  
...  
Keyword(s):  
Isotopic Labeling ◽  
Central Domain ◽  
Multidomain Protein ◽  
Segmental Isotopic Labeling

Segmental Isotopic Labeling of a Central Domain in a Multidomain Protein by ProteinTrans-Splicing Using Only One Robust DnaE Intein

2009 ◽  
Vol 121 (33) ◽  
pp. 6244-6247 ◽  
Author(s):  
Alena E. L. Busche ◽  
A. Sesilja Aranko ◽  
Mehdi Talebzadeh-Farooji ◽  
Frank Bernhard ◽  
Volker Dötsch ◽  
...  
Keyword(s):  
Isotopic Labeling ◽  
Central Domain ◽  
Multidomain Protein ◽  
Segmental Isotopic Labeling

Segmental isotopic labeling of multi-domain and fusion proteins by protein trans-splicing in vivo and in vitro

2010 ◽  
Vol 5 (3) ◽  
pp. 574-587 ◽  
Author(s):  
Mikko Muona ◽  
A Sesilja Aranko ◽  
Vytas Raulinaitis ◽  
Hideo Iwaï
Keyword(s):  
Fusion Proteins ◽  
Isotopic Labeling ◽  
Trans Splicing ◽  
Segmental Isotopic Labeling

Segmental Isotopic Labeling of Ubiquitin Chains To Unravel Monomer-Specific Molecular Behavior

2011 ◽  
Vol 123 (47) ◽  
pp. 11406-11410 ◽  
Author(s):  
Carlos A. Castañeda ◽  
Liat Spasser ◽  
Sudhir N. Bavikar ◽  
Ashraf Brik ◽  
David Fushman
Keyword(s):  
Isotopic Labeling ◽  
Molecular Behavior ◽  
Segmental Isotopic Labeling

scholarly journals Segmental isotopic labeling by asparaginyl endopeptidase-mediated protein ligation

2018 ◽  
Vol 71 (4) ◽  
pp. 225-235 ◽  
Author(s):  
Kornelia M. Mikula ◽  
Luisa Krumwiede ◽  
Andreas Plückthun ◽  
Hideo Iwaï
Keyword(s):  
Isotopic Labeling ◽  
Protein Ligation ◽  
Asparaginyl Endopeptidase ◽  
Segmental Isotopic Labeling

scholarly journals Optimization of sortase A ligation for flexible engineering of complex protein systems

2020 ◽  
Vol 295 (9) ◽  
pp. 2664-2675 ◽  
Author(s):  
Jess Li ◽  
Yue Zhang ◽  
Olivier Soubias ◽  
Domarin Khago ◽  
Fa-an Chao ◽  
...  
Keyword(s):  
Posttranslational Modifications ◽  
High Efficiency ◽  
Isotopic Labeling ◽  
Side Reactions ◽  
Sortase A ◽  
Chemical Ligation ◽  
Multidomain Protein ◽  
Domain Specific ◽  
Wide Range ◽  
Complex Protein

Engineering and bioconjugation of proteins is a critically valuable tool that can facilitate a wide range of biophysical and structural studies. The ability to orthogonally tag or label a domain within a multidomain protein may be complicated by undesirable side reactions to noninvolved domains. Furthermore, the advantages of segmental (or domain-specific) isotopic labeling for NMR, or deuteration for neutron scattering or diffraction, can be realized by an efficient ligation procedure. Common methods—expressed protein ligation, protein trans-splicing, and native chemical ligation—each have specific limitations. Here, we evaluated the use of different variants of Staphylococcus aureus sortase A for a range of ligation reactions and demonstrate that conditions can readily be optimized to yield high efficiency (i.e. completeness of ligation), ease of purification, and functionality in detergents. These properties may enable joining of single domains into multidomain proteins, lipidation to mimic posttranslational modifications, and formation of cyclic proteins to aid in the development of nanodisc membrane mimetics. We anticipate that the method for ligating separate domains into a single functional multidomain protein reported here may enable many applications in structural biology.

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