Properties of Mutant Photosynthetic Reaction Centers of Purple Non-Sulfur Bacteria Cereibacter sphaeroides with M206 Ile→Gln Substitution

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Tatiana Yu. Fufina¹, Olga A. Tretchikova¹, Anton M. Khristin¹, Ravil A. Khatypov¹, and Lyudmila G. Vasilieva^1,a*

¹Institute of Basic Biological Problems, Pushchino Scientific Center for Biological Research, Russian Academy of Sciences, 142290 Pushchino, Moscow Region, Russia

^* To whom correspondence should be addressed.

Received June 9, 2022; Revised August 14, 2022; Accepted August 14, 2022

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In the structure of photosynthetic reaction center (RC) of the purple bacterium Cereibacter sphaeroides the highly conserved amino acid residue Ile-M206 is located near the bacteriochlorophyll dimer P, which is the primary electron donor, and the monomeric bacteriochlorophyll B_A, which is the nearest electron acceptor. Since Ile-M206 is close to the C2-acetyl group of bacteriochlorophyll P_B, the hydroxyl group of Tyr-M210, and to the C9-keto group of bacteriochlorophyll B_A, as well as to the water molecule near the latter group, this site can be used for introducing mutations in order to study mechanisms of primary photochemical processes in the RC. Previously it was shown that the Ile→Glu substitution at the M204 position (analog of M206 in the RC of C. sphaeroides) in the RC of the closely related purple non-sulfur bacterium Rhodobacter capsulatus significantly affected kinetics of the P⁺H_A^– state formation, whereas the M204 Ile→Gln substitution led to the loss of BChl B_A molecule from the complex structure. In the present work, it is shown that the single I(M206)Q or double I(M206)Q + F(M208)A amino acid substitutions in the RC of C. sphaeroides do not change the pigment composition and do not markedly influence redox potential of the primary electron donor. However, substitution of Ile M206 by Gln affected positions and amplitudes of the absorption bands of bacteriochlorophylls, increased lifetime of the primary electron donor P* excited state from 3.1 ps to 22 ps, and decreased quantum yield of the P⁺Q_A^– state formation to 60%. These data suggest significant changes in the pigment–protein interactions in the vicinity of the primary electron donor P and the nearest electron acceptor B_A. A considerable decrease was also noticed in the resistance of the mutant RC to thermal denaturation, which was more pronounced in the RC with the double substitution I(M206)Q + F(M208)A. This was likely associated with the disruption of the dense packing of the protein near bacteriochlorophylls P_B and B_A. Possible reasons for different effects of identical mutations on the properties of two highly homologous RCs from closely related purple non-sulfur bacteria are discussed.

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KEY WORDS: photosynthesis reaction center, Rhodobacter capsulatus, Cereibacter sphaeroides, bacteriochlorophyll, purple non-sulfur bacteria, photochemical charge separation, quantum yield, pigment composition, redox potential, primary electron donor, thermal stability of membrane proteins

DOI: 10.1134/S000629792210008X

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