\u3csup\u3e1\u3c/sup\u3eH, \u3csup\u3e13\u3c/sup\u3eC, and \u3csup\u3e15\u3c/sup\u3eN assignments for the \u3ci\u3eArchaeglobus fulgidis \u3c/i\u3eprotein AF2095

Structural genomics is providing a means to determine the molecular and cellular function for the vast amount of proteins in the Human proteome that lack any explicit experimental information by characterizing the complete range of protein folds (Montelione, 2001). The Northeast Structural Genomics Consortium (NESG; http://www.nesg.org/) is a pilot project funded by the National Institutes of Health Protein Structure Initiative, focusing on proteins from eukaryotic model organisms including humans. The thermophillic archaea Archaeglobus fulgidis AF2095 protein is an example of a protein of unknown biological function targeted for structural analysis by NESG. AF2095 belongs to the Pfam family PF01981 – UPF0099, protein domain family of unknown function that has been found in yeast, archaebacteria and eubacteria. AF2095 has been assigned to NESG Cluster ID:17431, a set of fourteen proteins with high (\u3e~30%) sequence identity with human, Drosophila, Caenorhabditis elegans, Arabidopsis, yeast, archaeal and eubacterial origin (Liu, 2004). A total of fifty-six proteins are identified when the analysis is expanded to include all available genomes, where determining the NMR solution structure of AF2095 can be leveraged to infer 3D structural information for these proteins. Here we report the near complete 1H, 15N, and 13C NMR as signments and secondary structure of AF2095. These data provide a basis for determining the solution structure of AF2095, for further investigation of the func tion of this protein and for providing representative structural and functional information for the protein domain family that includes AF2095. This document includes the supplementary material originally published online only

Solution structure of \u3ci\u3eArchaeglobus fulgidis\u3c/i\u3e peptidyl-tRNA hydrolase (Pth2) provides evidence for an extensive conserved family of Pth2 enzymes in archea, bacteria, and eukaryotes

The solution structure of protein AF2095 from the thermophilic archaea Archaeglobus fulgidis, a 123- residue (13.6-kDa) protein, has been determined by NMR methods. The structure of AF2095 is comprised of four α-helices and a mixed β-sheet consisting of four parallel and anti-parallel β-strands, where the α-helices sandwich the β-sheet. Sequence and structural comparison of AF2095 with proteins from Homo sapiens, Methanocaldococcus jannaschii, and Sulfolobus solfataricus reveals that AF2095 is a peptidyltRNA hydrolase (Pth2). This structural comparison also identifies putative catalytic residues and a tRNA interaction region for AF2095. The structure of AF2095 is also similar to the structure of protein TA0108 from archaea Thermoplasma acidophilum, which is deposited in the Protein Data Bank but not functionally annotated. The NMR structure of AF2095 has been further leveraged to obtain good-quality structural models for 55 other proteins. Although earlier studies have proposed that the Pth2 protein family is restricted to archeal and eukaryotic organisms, the similarity of the AF2095 structure to human Pth2, the conservation of key active-site residues, and the good quality of the resulting homology models demonstrate a large family of homologous Pth2 proteins that are conserved in eukaryotic, archaeal, and bacterial organisms, providing novel insights in the evolution of the Pth and Pth2 enzyme families

Solution structure of Archaeglobus fulgidis peptidyl-tRNA hydrolase (Pth2) provides evidence for an extensive conserved family of Pth2 enzymes in archea, bacteria, and eukaryotes

The solution structure of protein AF2095 from the thermophilic archaea Archaeglobus fulgidis, a 123-residue (13.6-kDa) protein, has been determined by NMR methods. The structure of AF2095 is comprised of four α-helices and a mixed β-sheet consisting of four parallel and anti-parallel β-strands, where the α-helices sandwich the β-sheet. Sequence and structural comparison of AF2095 with proteins from Homo sapiens, Methanocaldococcus jannaschii, and Sulfolobus solfataricus reveals that AF2095 is a peptidyl-tRNA hydrolase (Pth2). This structural comparison also identifies putative catalytic residues and a tRNA interaction region for AF2095. The structure of AF2095 is also similar to the structure of protein TA0108 from archaea Thermoplasma acidophilum, which is deposited in the Protein Data Bank but not functionally annotated. The NMR structure of AF2095 has been further leveraged to obtain good-quality structural models for 55 other proteins. Although earlier studies have proposed that the Pth2 protein family is restricted to archeal and eukaryotic organisms, the similarity of the AF2095 structure to human Pth2, the conservation of key active-site residues, and the good quality of the resulting homology models demonstrate a large family of homologous Pth2 proteins that are conserved in eukaryotic, archaeal, and bacterial organisms, providing novel insights in the evolution of the Pth and Pth2 enzyme families