Conformational dynamics of the transcriptional regulator CooA protein studied by subpicosecond mid-infrared vibrational spectroscopy.
CooA, which is a transcriptional regulator heme protein allosterically triggered by CO, is studied by femtosecond visible-pump mid-IR-probe spectroscopy. Transient bleaching upon excitation of the heme in the Soret band is detected at approximately 1979 cm(-1), which is the absorption region of the CO bound to the heme. The bleach signal shows a nonexponential decay with time constants of 56 and 290 ps, caused by the rebinding of the CO to the heme. About 98% of dissociated CO recombines geminately. The geminate recombination rate in CooA is significantly faster than those in myoglobin and hemoglobin. The angle of the bound CO with respect to the porphyrin plane is calculated to be about 78 degrees on the basis of the anisotropy measurements. A shift of the bleached mid-IR spectrum of the bound CO is detected and has a characteristic time of 160 ps. It is suggested that the spectral shift is caused by a difference in the frequency of the bound CO in different protein conformations, particularly in an active conformation and in an intermediate one, which is on the way toward an inactive conformation. Thus, the biologically relevant conformation change in CooA was traced. Possible assignment of the observed conformation change is discussed.
Rubtsov
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School of Materials Science, Japan Advanced Institute of Science and Technology, Tatsunokuchi, Ishikawa 923-1292, Japan. irubtsov@sas.upenn.edu
Zhang
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T
Nakajima
H
H
Aono
S
S
Rubtsov
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GI
Kumazaki
S
S
Yoshihara
K
K
D001426:Bacterial Proteins; D002248:Carbon Monoxide / Q000737:chemistry; D006418:Heme / Q000737:chemistry; D006420:Hemeproteins / Q000737:chemistry*; D006454:Hemoglobins / Q000737:chemistry; D007700:Kinetics; D009211:Myoglobin / Q000737:chemistry; D011487:Protein Conformation; D013055:Spectrophotometry, Infrared / Q000379:methods; D015534:Trans-Activators / Q000737:chemistry*
D001426:Bacterial Proteins; C108776:CooA protein, Rhodospirillum rubrum; D006420:Hemeproteins; D006454:Hemoglobins; D009211:Myoglobin; D015534:Trans-Activators; D006418:Heme; D002248:Carbon Monoxide