Arrangement of RecA protein in its active filament determined by polarized-light spectroscopy.
Linear dichroism (LD) polarized-light spectroscopy is used to determine the arrangement of RecA in its large filamentous complex with DNA, active in homologous recombination. Angular orientation data for two tryptophan and seven tyrosine residues, deduced from differential LD of wild-type RecA vs. mutants that were engineered to attenuate the UV absorption of selected residues, revealed a rotation by some 40 degrees of the RecA subunits relative to the arrangement in crystal without DNA. In addition, conformational changes are observed for tyrosine residues assigned to be involved in DNA binding and in RecA-RecA contacts, thus potentially related to the global structure of the filament and its biological function. The presented spectroscopic approach, called "Site-Specific Linear Dichroism" (SSLD), may find forceful applications also to other biologically important fibrous complexes not amenable to x-ray crystallographic or NMR structural analysis.
Department of Physical Chemistry, Chalmers University of Technology, S-41296 Gothenburg, Sweden.
Katsumi
K
Morimatsu
Masayuki
M
Takahashi
Bengt
B
Nordén
D008027:Light; D011487:Protein Conformation; D011938:Rec A Recombinases / Q000737:chemistry*; D013057:Spectrum Analysis / Q000379:methods*; D014364:Tryptophan / Q000737:chemistry; D014443:Tyrosine / Q000737:chemistry
D014443:Tyrosine; D014364:Tryptophan; D011938:Rec A Recombinases