Faculty Collaboration Database

A hybrid structural model of the complete Brugia malayi cytoplasmic asparaginyl-tRNA synthetase. J Mol Biol 2011 Jan 28;405(4):1056-69

Date

12/08/2010

Pubmed ID

21134380

DOI

10.1016/j.jmb.2010.11.049

Scopus ID

2-s2.0-79251595309 (requires institutional sign-in at Scopus site)   35 Citations

Abstract

Aminoacyl-tRNA synthetases are validated molecular targets for anti-infective drug discovery because of their essentiality in protein synthesis. Thanks to genome sequencing, it is now possible to systematically study aminoacyl-tRNA synthetases from human eukaryotic parasites as putative targets for novel drug discovery. As part of a program targeting class IIb asparaginyl-tRNA synthetases (AsnRS) from the parasitic nematode Brugia malayi for anti-filarial drugs, we report the complete structure of a eukaryotic AsnRS. Metazoan and fungal AsnRS differ from their bacterial homologues by the addition of a conserved N-terminal extension of about 110 residues whose structure we have determined by solution NMR for the B. malayi enzyme. In addition, we solved by X-ray crystallography a series of structures of the catalytically active N-terminally truncated enzyme (residues 112-548), allowing the structural basis for the mechanism of asparagine activation to be elucidated. The N-terminal domain contains a structured region with a novel fold featuring a lysine-rich helix that is shown by NMR to interact with tRNA. This is connected by an unstructured tether to the remainder of the enzyme, which is highly similar to the known structure of bacterial AsnRS. These data enable a model of the complete AsnRS-tRNA complex to be constructed.

Author List

Crepin T, Peterson F, Haertlein M, Jensen D, Wang C, Cusack S, Kron M

Authors

Michael Kron MD Director, Professor in the Medicine department at Medical College of Wisconsin
Francis C. Peterson PhD Professor in the Biochemistry department at Medical College of Wisconsin

MESH terms used to index this publication - Major topics in bold

Amino Acid Sequence
Animals
Aspartate-tRNA Ligase
Base Sequence
Brugia malayi
Catalytic Domain
Crystallography, X-Ray
Cytoplasm
DNA Primers
Enzyme Activation
Helminth Proteins
Humans
Models, Molecular
Molecular Sequence Data
Nuclear Magnetic Resonance, Biomolecular
Protein Structure, Tertiary
RNA, Transfer, Amino Acyl
Recombinant Proteins
Sequence Homology, Amino Acid