A time-resolved resonance Raman analysis shows that the vinyl stretching band (1629 cm
−1) of the O
2-binding heme of cytochrome
c oxidase shifts to 1627 cm
−1 instantaneously upon photolysis of CO and remains for at least 5 ms before reaching the static band (1626 cm
−1). Within the same time scale, an intermediate vinyl bending mode of another heme appears at 435 cm
−1. These results suggest that both hemes cooperatively control O
2 binding by forming an intermediate conformation for effective proton pumping.
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