Evidence for displacements of the C-helix by CO ligation and DNA binding to CooA revealed by UV resonance Raman spectroscopy.
The UV and visible resonance Raman spectra are reported for CooA from Rhodospirillum rubrum, which is a transcriptional regulator activated by growth in a CO atmosphere. CO binding to heme in its sensor domain causes rearrangement of its DNA-binding domain, allowing binding of DNA with a specific sequence. The sensor and DNA-binding domains are linked by a hinge region that follows a long C-helix. UV resonance Raman bands arising from Trp-110 in the C-helix revealed local movement around Trp-110 upon CO binding. The indole side chain of Trp-110, which is exposed to solvent in the CO-free ferrous state, becomes buried in the CO-bound state with a slight change in its orientation but maintains a hydrogen bond with a water molecule at the indole nitrogen. This is the first experimental data supporting a previously proposed model involving displacement of the C-helix and heme sliding. The UV resonance Raman spectra for the CooA-DNA complex indicated that binding of DNA to CooA induces a further displacement of the C-helix in the same direction during transition to the complete active conformation. The Fe-CO and C-O stretching bands showed frequency shifts upon DNA binding, but the Fe-His stretching band did not. Moreover, CO-geminate recombination was more efficient in the DNA-bound state. These results suggest that the C-helix displacement in the DNA-bound form causes the CO binding pocket to narrow and become more negative.
Kubo
Minoru
M
Okazaki Institute for Integrative Bioscience, National Institutes of Natural Sciences, Okazaki 444-8787, Japan.
Inagaki
Sayaka
S
Yoshioka
Shiro
S
Uchida
Takeshi
T
Mizutani
Yasuhisa
Y
Aono
Shigetoshi
S
Kitagawa
Teizo
T
D001426:Bacterial Proteins / Q000737:chemistry* / Q000378:metabolism*; D002248:Carbon Monoxide / Q000737:chemistry; D004247:DNA / Q000737:chemistry*; D004583:Electrons; D006418:Heme / Q000737:chemistry; D006420:Hemeproteins / Q000737:chemistry* / Q000378:metabolism*; D007501:Iron / Q000378:metabolism; D007700:Kinetics; D008024:Ligands; D008958:Models, Molecular; D008968:Molecular Conformation; D011485:Protein Binding; D011487:Protein Conformation; D017433:Protein Structure, Secondary; D012247:Rhodospirillum rubrum / Q000378:metabolism; D013056:Spectrophotometry, Ultraviolet / Q000379:methods*; D013059:Spectrum Analysis, Raman; D013997:Time Factors; D015534:Trans-Activators / Q000737:chemistry* / Q000378:metabolism*; D014364:Tryptophan / Q000737:chemistry; D014466:Ultraviolet Rays
D001426:Bacterial Proteins; C108776:CooA protein, Rhodospirillum rubrum; D006420:Hemeproteins; D008024:Ligands; D015534:Trans-Activators; D006418:Heme; D002248:Carbon Monoxide; D014364:Tryptophan; D004247:DNA; D007501:Iron