TY - JOUR
T1 - Many combinations of amino acid sequences in a conserved region of the D1 protein satisfy photosystem II function
AU - Kless, Hadar
AU - Vermaas, Willem
N1 - Funding Information:
This research is supported by a grant from the National Science Foundation (DMB90-58279). This is publication no. 218 from the Arizona State University Center for the Study of Early Events in Photosynthesis.
PY - 1995
Y1 - 1995
N2 - The putative de helix of the D1 protein is located at the accepter side of photosystem II (PS II) and serves as an indispensable part of a niche that binds the secondary plastoquinone Q(B). Combinatorial mutagenesis was applied to a stretch of four residues in a highly conserved region of this putative helix in order to reveal amino acid combinations that are able to support PS II function. An obligate photoheteroirophic mutant of the cyanobacterium Synechocystis sp. PCC 6803, missing four residues (ΔYFGR254-7 in the de helix, was transformed with a D1-coding sequence carrying fully degenerate combinations of codons at the site of the deletion. Upon selection for photoautotrophy, 25 mutants with functional PS II were isolated. All mutants showed different codon combinations at positions 254 to 257; none was identical to the wild-type sequence, and none of the conserved residues was found to be mandatory for PS II function. However, 24 of the mutants contained Tyr or Phe at position 254 while at the other three positions many different amino acid combinations could be functionally accommodated. Most sequences maintained an amphiphilic arrangement of the helix that may align Tyr254 facing the Q(B) binding pocket. This residue is proposed to be functionally analogous to Phe216 of the L subunit in purple bacteria which contributes to binding of Q(B). Most of the PS II properties were similar in the mutants compared to wild-type. Noticeable modifications in the mutants concerned the semiquinone equilibrium of electron transfer between Q(A) and Q(B), and the affinity of PS II inhibitors. Differential effects on the semiquinone equilibrium were observed between two distinct quinones occupying the Q(B) Site (plastoquinone versus 2,5-dichloro-p-benzoquinone), implying that residues in this domain are involved, directly or indirectly, with different binding determinants of the quinones. Even though many different combinations of amino acids in positions 254 to 257 of the D1 protein may satisfy the primary function of PS II, complex requirements need to be combined for optimized performance of the Q(B) binding niche.
AB - The putative de helix of the D1 protein is located at the accepter side of photosystem II (PS II) and serves as an indispensable part of a niche that binds the secondary plastoquinone Q(B). Combinatorial mutagenesis was applied to a stretch of four residues in a highly conserved region of this putative helix in order to reveal amino acid combinations that are able to support PS II function. An obligate photoheteroirophic mutant of the cyanobacterium Synechocystis sp. PCC 6803, missing four residues (ΔYFGR254-7 in the de helix, was transformed with a D1-coding sequence carrying fully degenerate combinations of codons at the site of the deletion. Upon selection for photoautotrophy, 25 mutants with functional PS II were isolated. All mutants showed different codon combinations at positions 254 to 257; none was identical to the wild-type sequence, and none of the conserved residues was found to be mandatory for PS II function. However, 24 of the mutants contained Tyr or Phe at position 254 while at the other three positions many different amino acid combinations could be functionally accommodated. Most sequences maintained an amphiphilic arrangement of the helix that may align Tyr254 facing the Q(B) binding pocket. This residue is proposed to be functionally analogous to Phe216 of the L subunit in purple bacteria which contributes to binding of Q(B). Most of the PS II properties were similar in the mutants compared to wild-type. Noticeable modifications in the mutants concerned the semiquinone equilibrium of electron transfer between Q(A) and Q(B), and the affinity of PS II inhibitors. Differential effects on the semiquinone equilibrium were observed between two distinct quinones occupying the Q(B) Site (plastoquinone versus 2,5-dichloro-p-benzoquinone), implying that residues in this domain are involved, directly or indirectly, with different binding determinants of the quinones. Even though many different combinations of amino acids in positions 254 to 257 of the D1 protein may satisfy the primary function of PS II, complex requirements need to be combined for optimized performance of the Q(B) binding niche.
KW - Combinatorial mutagenesis
KW - Cyanobacteria
KW - Molecular evolution
KW - Protein-cofactor interaction
KW - Reaction center
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U2 - 10.1006/jmbi.1994.0071
DO - 10.1006/jmbi.1994.0071
M3 - Article
C2 - 7853392
AN - SCOPUS:0028902008
VL - 246
SP - 120
EP - 131
JO - Journal of Molecular Biology
JF - Journal of Molecular Biology
SN - 0022-2836
IS - 6
ER -