Showing papers by "Gautam Basu published in 2009"
TL;DR: The results point towards conformational heterogeneity of water‐soluble Aib‐based peptide helices and the associated subtleties.
Abstract: The amino acid Aib predisposes a peptide to be helical with context-dependent preference for either 3(10)- or alpha- or a mixed helical conformation. Short peptides also show an inherent tendency to be unfolded. To characterize helical and unfolded states adopted by water-soluble Aib-containing peptides, the conformational preference of Ac-Ala-Aib-Ala-Lys-Ala-Aib-Lys-Ala-Lys-Ala-Aib-Tyr-NH(2) was determined by CD, NMR and MD simulations as a function of temperature. Temperature-dependent CD data indicated the contribution of two major components, each an admixture of helical and extended/polyproline II structures. Both right- and left-handed helical conformations were detected from deconvolution of CD data and (13)C NMR experiments. The presence of a helical backbone, more pronounced at the N-terminal, and a temperature-induced shift in alpha-helix/3(10)-helix equilibrium, more pronounced at the C-terminal, emerged from NMR data. Starting from polyproline II, the N-terminal of the peptide folded into a helical backbone in MD simulations within 5 ns at 60 degrees C. Longer simulations showed a mixed-helical backbone to be stable over the entire peptide at 5 degrees C while at 60 degrees C the mixed-helix was either stable at the N-terminus or occurred in short stretches through out the peptide, along with a significant population of polyproline II. Our results point towards conformational heterogeneity of water-soluble Aib-based peptide helices and the associated subtleties. The problem of analyzing CD and NMR data of both left- and right-handed helices are discussed, especially the validity of the ellipticity ratio [theta](222)/[theta](207), as a reporter of alpha-/3(10)- population ratio, in right- and left-handed helical mixtures.
29 citations
TL;DR: It is shown that an extended loop, present in the anticodon-binding domains of GlnRSs, is absent in archaeal GluRS, suggesting that the loop was a later addition, generating additional anti-codon discrimination capability in Gln RS as it evolved from Glu RS in eukaryotes.
Abstract: aaRSs (aminoacyl-tRNA synthetases) are multi-domain proteins that have evolved by domain acquisition. The anti-codon binding domain was added to the more ancient catalytic domain during aaRS evolution. Unlike in eukaryotes, the anti-codon binding domains of GluRS (glutamyl-tRNA synthetase) and GlnRS (glutaminyl-tRNA synthetase) in bacteria are structurally distinct. This originates from the unique evolutionary history of GlnRSs. Starting from the catalytic domain, eukaryotic GluRS evolved by acquiring the archaea/eukaryote-specific anti-codon binding domain after branching away from the eubacteria family. Subsequently, eukaryotic GlnRS evolved from GluRS by gene duplication and horizontally transferred to bacteria. In order to study the properties of the putative ancestral GluRS in eukaryotes, formed immediately after acquiring the anti-codon binding domain, we have designed and constructed a chimaeric protein, cGluGlnRS, consisting of the catalytic domain, Ec GluRS (Escherichia coli GluRS), and the anti-codon binding domain of EcGlnRS (E. coli GlnRS). In contrast to the isolated EcN-GluRS, cGluGlnRS showed detectable activity of glutamylation of E. coli tRNA(glu) and was capable of complementing an E. coli ts (temperature-sensitive)-GluRS strain at non-permissive temperatures. Both cGluGlnRS and EcN-GluRS were found to bind E. coli tRNA(glu) with native EcGluRS-like affinity, suggesting that the anticodon-binding domain in cGluGlnRS enhances k(cat) for glutamylation. This was further confirmed from similar experiments with a chimaera between EcN-GluRS and the substrate-binding domain of EcDnaK (E. coli DnaK). We also show that an extended loop, present in the anticodon-binding domains of GlnRSs, is absent in archaeal GluRS, suggesting that the loop was a later addition, generating additional anti-codon discrimination capability in GlnRS as it evolved from GluRS in eukaryotes.
13 citations
TL;DR: It is demonstrated that in addition to the anticodon‐binding domain, tRNAGln discriminatory elements may be present in the catalytic domain in E. coli GluRS as well.
11 citations
TL;DR: From an analysis of Coulomb energy distributions of a large set of protein-protein complexes, it is shown that the positive tail in the energy distribution disappears when the monopole-monopole term, the only energy term independent of inter-subunit orientations, is removed, indicating that unfavorable Coulomb energies associated with subunit orientation are excluded in protein- protein complexes.
Abstract: From an analysis of Coulomb energy distributions of a large set of protein–protein complexes we show that the positive tail in the energy distribution disappears when the monopole–monopole term, the only energy term independent of inter-subunit orientations, is removed. This indicates that unfavorable Coulomb energies associated with subunit orientations are excluded in protein–protein complexes. The overall result remained unchanged when solvent effects were included. Our results have important bearing on the restriction of subunit orientations in protein–protein complexes and complement a recent work [K. Brock, K. Talley, K. Coley, P. Kundrotas, E. Alexov, Optimization of electrostatic interactions in protein–protein complexes, Biophys. J. 93 (2007) 3340–3352.] which showed that Coulomb energy of interaction in protein–protein complexes is sequence-optimized.
5 citations