Probing the environment of cu(b) in heme-copper oxidases.
Time-resolved step-scan FTIR (TRS2-FTIR) and density functional theory have been applied to probe the structural dynamics of CuB in heme-copper oxidases at room temperature. The TRS2-FTIR data of cbb3 from Pseudomonas stutzeri indicate a small variation in the frequency of the transient CO bound to CuB in the pH/pD 7-9 range. This observation in conjunction with density functional theory calculations, in which significant frequency shifts of the nu(CO) are observed upon deprotonation and/or detachment of the CuB ligands, demonstrates that the properties of the CuB ligands including the cross-linked tyrosine, in contrast to previous reports, remain unchanged in the pH 7-9 range. We attribute the small variations in the nu(CO) of CuB to protein conformational changes in the vicinity of CuB. Consequently, the split of the heme Fe-CO vibrations (alpha-, beta-, and gamma-forms) is not due to changes in the ligation and/or protonation states of the CuB ligands or to the presence of one or more ionizable groups, as previously suggested, but the result of global protein conformational changes in the vicinity of CuB which, in turn, affect the position of CuB with respect to the heme Fe.
Daskalakis
Vangelis
V
Department of Chemistry, University of Crete, 71003 Voutes, Heraklion, Crete, Greece.
Pinakoulaki
Eftychia
E
Stavrakis
Stavros
S
Varotsis
Constantinos
C
D001665:Binding Sites; D002248:Carbon Monoxide / Q000737:chemistry; D003300:Copper / Q000737:chemistry* / Q000378:metabolism; D003576:Electron Transport Complex IV / Q000737:chemistry* / Q000378:metabolism; D006418:Heme / Q000737:chemistry* / Q000378:metabolism; D008956:Models, Chemical; D008958:Models, Molecular; D009532:Nickel / Q000737:chemistry* / Q000378:metabolism; D010782:Photolysis; D044223:Pseudomonas stutzeri / Q000201:enzymology; D017550:Spectroscopy, Fourier Transform Infrared
C062136:nickel copper alloy; D006418:Heme; D003300:Copper; D009532:Nickel; D002248:Carbon Monoxide; C414443:cbb3 oxidase; D003576:Electron Transport Complex IV