TY - CHAP
T1 - Segmental isotopic labeling of proteins for NMR study using intein technology
AU - Liu, Dongsheng
AU - Cowburn, David
N1 - Publisher Copyright:
© Springer Science+Business Media New York 2017.
PY - 2017
Y1 - 2017
N2 - Segmental isotopic labeling of samples for NMR studies is attractive for large complex biomacromolecular systems, especially for studies of function-related protein-ligand interactions and protein dynamics (Goto and Kay, Curr Opin Struct Biol 10:585-592, 2000; Rosa et al., Molecules (Basel, Switzerland) 18:440, 2013; Hiroaki, Expert Opin Drug Discovery 8:523-536, 2013). Advantages of segmental isotopic labeling include selective examination of specific segment(s) within a protein by NMR, significantly reducing the spectral complexity for large proteins, and allowing for the application of a variety of solution-based NMR strategies. By utilizing intein techniques (Wood and Camarero, J Biol Chem 289:14512-14519, 2014; Paulus, Annu Rev Biochem 69:447-496, 2000), two related approaches can generally be used in the segmental isotopic labeling of proteins: expressed protein ligation (Muir, Annu Rev Biochem 72:249- 289, 2003) and protein trans -splicing (Shah et al., J Am Chem Soc 134:11338-11341, 2012). Here, we describe general implementation and latest improvements of expressed protein ligation method for the production of segmental isotopic labeled NMR samples.
AB - Segmental isotopic labeling of samples for NMR studies is attractive for large complex biomacromolecular systems, especially for studies of function-related protein-ligand interactions and protein dynamics (Goto and Kay, Curr Opin Struct Biol 10:585-592, 2000; Rosa et al., Molecules (Basel, Switzerland) 18:440, 2013; Hiroaki, Expert Opin Drug Discovery 8:523-536, 2013). Advantages of segmental isotopic labeling include selective examination of specific segment(s) within a protein by NMR, significantly reducing the spectral complexity for large proteins, and allowing for the application of a variety of solution-based NMR strategies. By utilizing intein techniques (Wood and Camarero, J Biol Chem 289:14512-14519, 2014; Paulus, Annu Rev Biochem 69:447-496, 2000), two related approaches can generally be used in the segmental isotopic labeling of proteins: expressed protein ligation (Muir, Annu Rev Biochem 72:249- 289, 2003) and protein trans -splicing (Shah et al., J Am Chem Soc 134:11338-11341, 2012). Here, we describe general implementation and latest improvements of expressed protein ligation method for the production of segmental isotopic labeled NMR samples.
KW - Expressed protein ligation
KW - Intein
KW - Isotopic labeling
KW - NMR
KW - Structural biology
UR - http://www.scopus.com/inward/record.url?scp=84991240706&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=84991240706&partnerID=8YFLogxK
U2 - 10.1007/978-1-4939-6451-2_9
DO - 10.1007/978-1-4939-6451-2_9
M3 - Chapter
C2 - 27714614
AN - SCOPUS:84991240706
T3 - Methods in Molecular Biology
SP - 131
EP - 145
BT - Methods in Molecular Biology
PB - Humana Press Inc.
ER -