Conformational change of the N-domain on formation of the complex between the GTPase domains of Thermus aquaticus Ffh and FtsY

Biochim Biophys Acta. 2002 May 20;1597(1):107-14. doi: 10.1016/s0167-4838(02)00287-x.

Abstract

The structural basis for the GTP-dependent co-translational targeting complex between the signal recognition particle (SRP) and its receptor is unknown. The complex has been shown to have unusual kinetics of formation, and association in vivo is likely to be dependent on catalysis by the SRP RNA. We have determined conditions for RNA-independent association of the 'NG' GTPase domains of the prokaryotic homologs of the SRP components, Ffh and FtsY, from Thermus aquaticus. Consistent with previous studies of the Escherichia coli proteins, the kinetics of association and dissociation are slow. The T. aquaticus FtsY is sensitive to an endogenous proteolytic activity that cleaves at two sites--the first in a lengthy linker peptide that spans the interface between the N and G domains, and the second near the N-terminus of the N domain of FtsY. Remarkably, this second cleavage occurs only on formation of the Ffh/FtsY complex. The change in protease sensitivity of this region, which is relatively unstructured in the FtsY but not in the Ffh NG domain, implies that it undergoes conformational change on formation of the complex between the two proteins. The N domain, therefore, participates in the interactions that mediate the GTP-dependent formation of the targeting complex.

Publication types

  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Amino Acid Sequence
  • Bacterial Proteins / chemistry
  • Bacterial Proteins / genetics
  • Bacterial Proteins / metabolism
  • Binding Sites
  • GTP Phosphohydrolases / chemistry*
  • Kinetics
  • Models, Molecular
  • Mutagenesis, Site-Directed
  • Peptide Hydrolases / chemistry
  • Protein Conformation
  • Protein Structure, Secondary
  • Receptors, Cytoplasmic and Nuclear / chemistry
  • Receptors, Cytoplasmic and Nuclear / genetics
  • Receptors, Cytoplasmic and Nuclear / metabolism*
  • Receptors, Peptide / chemistry
  • Receptors, Peptide / metabolism*
  • Signal Recognition Particle / chemistry
  • Signal Recognition Particle / metabolism*
  • Thermus / enzymology
  • Thermus / metabolism*

Substances

  • Bacterial Proteins
  • FtsY protein, Bacteria
  • Receptors, Cytoplasmic and Nuclear
  • Receptors, Peptide
  • Signal Recognition Particle
  • signal peptide receptor
  • Peptide Hydrolases
  • GTP Phosphohydrolases