title: FTIR spectroscopy of synechocystis 6803 mutants affected on the hydrogen bonds to the carbonyl groups of the PsaA chlorophyll of P700 supports an extensive delocalization of the charge in P700+.
abstract: P700, the primary electron donor of photosystem I is an asymmetric dimer made of one molecule of chlorophyll a' (P(A)) and one of chlorophyll a (P(B)). While the carbonyl groups of P(A) are involved in hydrogen-bonding interactions with several surrounding amino acid side chains and a water molecule, P(B) does not engage in hydrogen bonding with the protein. Light-induced FTIR difference spectroscopy of the photooxidation of P700 has been combined with site-directed mutagenesis in Synechocystis sp. PCC 6803 to investigate the influence of these hydrogen bonds on the structure of P700 and P700(+). When the residue Thr A739, which donates a hydrogen bond to the 9-keto C=O group of P(A), is changed to Phe, a differential signal at 1653(+)/1638(-) cm(-1) in the P700(+)/P700 FTIR difference spectrum upshifts by approximately 30-40 cm(-1), as expected for the rupture of the hydrogen bond or, at least, a strong decrease of its strength. The same upshift is also observed in the FTIR spectrum of a triple mutant in which the residues involved in the three main hydrogen bonds to the 9-keto and 10a-carbomethoxy groups of P(A) have been changed to the symmetry-related side chains present around P(B). In addition, the spectrum of the triple mutant shows a decrease of a differential signal around 1735 cm(-1) and the appearance of a new signal around 1760 cm(-1). This is consistent with the perturbation of a bound 10a-ester C=O group that becomes free in the triple mutant. All of these observations support the assignment scheme proposed previously for the carbonyls of P700 and P700(+) [Breton, J., Nabedryk, E., and Leibl, W. (1999) Biochemistry 38, 11585-11592] and therefore reinforce our previous conclusions that the positive charge in P700(+) is largely delocalized over P(A) and P(B).
authors.affiliation: Department of Biochemistry, Biophysics, and Molecular Biology, Iowa State University, Ames, Iowa 50011, USA.
authors.forename: Maria
authors.initials: M
authors.lastname: Pantelidou
authors.forename: Parag R
authors.initials: PR
authors.lastname: Chitnis
authors.forename: Jacques
authors.initials: J
authors.lastname: Breton
mesh_terms: D018829:Adhesins, Bacterial; D001426:Bacterial Proteins; D002244:Carbon / Q000737:chemistry; D002352:Carrier Proteins / Q000737:chemistry*; D002455:Cell Division; D002734:Chlorophyll / Q000737:chemistry*; D000458:Cyanobacteria / Q000378:metabolism*; D006860:Hydrogen Bonding; D008027:Light; D008074:Lipoproteins / Q000737:chemistry*; D026901:Membrane Transport Proteins; D008958:Models, Molecular; D016297:Mutagenesis, Site-Directed; D009154:Mutation; D009584:Nitrogen / Q000737:chemistry; D017550:Spectroscopy, Fourier Transform Infrared / Q000379:methods*; D013912:Threonine / Q000737:chemistry
chemical_list: D018829:Adhesins, Bacterial; D001426:Bacterial Proteins; D002352:Carrier Proteins; D008074:Lipoproteins; D026901:Membrane Transport Proteins; C085000:PsaA protein, Streptococcus; D002734:Chlorophyll; D013912:Threonine; D002244:Carbon; D009584:Nitrogen