Effects of hydrogen bonding to a bacteriochlorophyll-bacteriopheophytin dimer in reaction centers from Rhodobacter sphaeroides.
The properties of the primary electron donor in reaction centers from Rhodobacter sphaeroides have been investigated in mutants containing a bacteriochlorophyll (BChl)--bacteriopheophytin (BPhe) dimer with and without hydrogen bonds to the conjugated carbonyl groups. The heterodimer mutation His M202 to Leu was combined with each of the following mutations: His L168 to Phe, which should remove an existing hydrogen bond to the BChl molecule; Leu L131 to His, which should add a hydrogen bond to the BChl molecule; and Leu M160 to His and Phe M197 to His, each of which should add a hydrogen bond to the BPhe molecule [Rautter, J., Lendzian, F., Schulz, C., Fetsch, A., Kuhn M., Lin, X., Williams, J. C., Allen J. P., & Lubitz, W. (1995) Biochemistry 34, 8130-8143]. Pigment extractions and Fourier transform Raman spectra confirm that all of the mutants contain a heterodimer. The bands in the resonance Raman spectra arising from the BPhe molecule, which is selectively enhanced, exhibit the shifts expected for the addition of a hydrogen bond to the 9-keto and 2-acetyl carbonyl groups. The oxidation--reduction midpoint potential of the donor is increased by approximately 85 mV by the addition of a hydrogen bond to the BChl molecule but is only increased by approximately 15 mV by the addition of a hydrogen bond to the BPhe molecule. An increase in the rate of charge recombination from the primary quinone is correlated with an increase in the midpoint potential. The yield of electron transfer to the primary quinone is 5-fold reduced for the mutants with a hydrogen bond to the BPhe molecule. Room- and low-temperature optical absorption spectra show small differences from the features that are typical for the heterodimer, except that a large increase in absorption is observed around 860-900 nm for the donor Qy band in the mutant that adds a hydrogen bond to the BChl molecule. The changes in the optical spectra and the yield of electron transfer are consistent with a model in which the addition of a hydrogen bond to the BChl molecule increases the energy of an internal charge transfer state while the addition to the BPhe molecule stabilizes this state. The results show that the properties of the heterodimer are different depending on which side is hydrogen-bonded and suggest that the hydrogen bonds alter the energy of the internal charge transfer state in a well-defined manner.
Allen
J P
JP
Department of Chemistry and Biochemistry, Arizona State University, Tempe 85287-1604, USA.
Artz
K
K
Lin
X
X
Williams
J C
JC
Ivancich
A
A
Albouy
D
D
Mattioli
T A
TA
Fetsch
A
A
Kuhn
M
M
Lubitz
W
W
D000595:Amino Acid Sequence; D001429:Bacteriochlorophylls / Q000737:chemistry* / Q000378:metabolism; D005583:Fourier Analysis; D006639:Histidine; D006860:Hydrogen Bonding; D007930:Leucine; D045342:Light-Harvesting Protein Complexes; D046911:Macromolecular Substances; D016297:Mutagenesis, Site-Directed; D010674:Pheophytins / Q000737:chemistry* / Q000378:metabolism; D045322:Photosynthetic Reaction Center Complex Proteins / Q000737:chemistry* / Q000378:metabolism; D017354:Point Mutation; D011994:Recombinant Proteins / Q000737:chemistry / Q000378:metabolism; D012242:Rhodobacter sphaeroides / Q000378:metabolism*; D013053:Spectrophotometry; D013059:Spectrum Analysis, Raman
D001429:Bacteriochlorophylls; D045342:Light-Harvesting Protein Complexes; D046911:Macromolecular Substances; D010674:Pheophytins; D045322:Photosynthetic Reaction Center Complex Proteins; D011994:Recombinant Proteins; C006678:bacteriopheophytin; D006639:Histidine; D007930:Leucine