Characterization of conformational changes coupled to ligand photodissociation from the heme binding domain of FixL Article

Mikšovská, J, Suquet, C, Satterlee, JD et al. (2005). Characterization of conformational changes coupled to ligand photodissociation from the heme binding domain of FixL . BIOCHEMISTRY, 44(30), 10028-10036. 10.1021/bi047369b

cited authors

  • Mikšovská, J; Suquet, C; Satterlee, JD; Larsen, RW

authors

abstract

  • Using transient absorption spectroscopy and photoacoustic calorimetry (PAC), we have characterized carbon monoxide photodissociation and rebinding to two forms of the heme domain of Bradyrhizobium japonicum FixL. Transient absorption results for the complete heme domain (FixL residues 140-270) and a truncated heme domain (missing 11 residues on the N-teminal end and 14 amino acid residues on the C-terminal end of the full length heme domain) show similar rates for ligand rebinding to the five-coordinate heme domain and the absence of any transient intermediate on a microsecond time scale. Results from PAC studies show that both the truncated and complete heme domains undergo a contraction upon ligand photolysis. In addition, CO photolysis from the complete heme domain gives rise to an intermediate with a lifetime of ∼150 ns which is absent in the truncated heme domain. We attribute the 150 ns phase to ligand release to the solvent which may be accelerated in the case of the truncated domain. The initial contraction is attributed to changes in the charge distribution due to reorganization of the surface salt bridge formed between GM82 and Arg227 or possibly to reorientation of Arg206. Changes in the charge distribution may play an important role in communication between the sensor domain and the regulatory domain and thus may be part of the signal transduction pathway. © 2005 American Chemical Society.

publication date

  • August 2, 2005

published in

Digital Object Identifier (DOI)

start page

  • 10028

end page

  • 10036

volume

  • 44

issue

  • 30