2.9 Å Crystal structure of ligand-free tryptophanyl-tRNA synthetase: Domain movements fragment the adenine nucleotide binding site

Valentin A. Ilyin, Brenda Temple, Mei Hu, Genpei Li, Yuhui Yin, Patrice Vachette, Charles W. Carter

Research output: Contribution to journalArticle

62 Scopus citations

Abstract

The crystal structure of ligand-free tryptophanyl-TRNA synthetase (TrpRS) was solved at 2.9 Å using a combination of molecular replacement and maximum-entropy map/phase improvement. The dimeric structure (R = 23.7, R(free) = 26.2) is asymmetric, unlike that of the TrpRS tryptophanyl-5'AMP complex (TAM; Doublie S, Bricogne G, Gilmore CJ, Carter CW Jr, 1995, Structure 3:17-31). In agreement with small-angle solution X-ray scattering experiments, unliganded TrpRS has a conformation in which both monomers open, leaving only the tryptophan-binding regions of their active gites intact. The amino terminal αA-helix, TIGN, and KMSKS signature sequences, and the distal helical domain rotate as a single rigid body away from the dinucleotide- binding fold domain, opening the AMP binding site, seen in the TAM complex, into two halves. Comparison of side-chain packing in ligand-free TrpRS and the TAM complex, using identification of nonpolar nuclei (Ilyin VA, 1994, Protein Eng 7:1189-1195), shows that significant repacking occurs between three relatively stable core regions, one of which acts as a bearing between the other two. These domain rearrangements provide a new structural paradigm that is consistent in detail with the 'induced-fit' mechanism proposed for TyrRS by Fersht et al. (Fersht AR, Knill-Jones JW, Beduelle H, Winter G, 1988, Biochemistry 27:1581-1587). Coupling of ATP binding determinants associated with the two catalytic signature sequences to the helical domain containing the presumptive anticodon-binding site provides a mechanism to coordinate active-site chemistry with relocation of the major tRNA binding determinants.

Original languageEnglish (US)
Pages (from-to)218-231
Number of pages14
JournalProtein Science
Volume9
Issue number2
DOIs
StatePublished - Jan 1 2000

Keywords

  • ATP binding site
  • Induced fit
  • Maximum entropy
  • Nonpolar nuclei
  • Tryptophanyl-tRNA synthetase

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology

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