Premium
The N‐terminal domain (IF2N) of bacterial translation initiation factor IF2 is connected to the conserved C‐terminal domains by a flexible linker
Author(s) -
Laursen Brian Søgaard,
Kjærgaard Anne Cecillie,
Mortensen Kim Kusk,
Hoffman David W.,
SperlingPetersen Hans Uffe
Publication year - 2004
Publication title -
protein science
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 3.353
H-Index - 175
eISSN - 1469-896X
pISSN - 0961-8368
DOI - 10.1110/ps.03337604
Subject(s) - linker , crystallography , eukaryotic translation , c terminus , translation (biology) , circular dichroism , biophysics , chemistry , biology , biochemistry , messenger rna , amino acid , gene , computer science , operating system
Bacterial translation initiation factor IF2 is a multidomain protein that is an essential component of a system for ensuring that protein synthesis begins at the correct codon within a messenger RNA. Full‐length IF2 from Escherichia coli and seven fragments of the protein were expressed, purified, and characterized using nuclear magnetic resonance (NMR) and circular dichroism (CD) methods. Interestingly, resonances of the 6 kD IF2N domain located at the extreme N terminus of IF2 can be clearly identified within the NMR spectra of the full‐length 97‐kD protein. 15 N NMR relaxation rate data indicate that (1) the IF2N domain is internally well ordered and tumbles in solution in a manner that is independent of the other domains of the IF2 protein, and (2) the IF2N domain is connected to the C‐terminal regions of IF2 by a flexible linker. Chemical shifts of resonances within the isolated IF2N domain do not significantly differ from those of the corresponding residues within the context of the full‐length 97‐kD protein, indicating that IF2N is a structurally independent unit that does not strongly interact with other regions of IF2. CD and NMR data together provide evidence that Domains I–III of IF2 have unstructured and flexible regions as well as substantial helical content; CD data indicate that the helical content of these regions decreases significantly at temperatures above 35°C. The features of structurally well‐ordered N‐ and C‐terminal domains connected by a flexible linker with significant helical content are reminiscent of another translation initiation factor, IF3.