2021
DOI: 10.26434/chemrxiv.14691339.v1
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Photosynthetic Reaction Center Variants Made via Genetic Code Expansion Show Tyr at M210 Tunes the Initial Electron Transfer Mechanism

Abstract: Photosynthetic reaction centers (RCs) from Rhodobacter sphaeroides were engineered to vary the electronic properties of a key tyrosine close to an essential electron transfer component (M210) via its replacement with site-specific genetically encoded noncanonical amino acid tyrosine analogs. High fidelity of noncanonical amino acid incorporation was verified with mass spectrometry and x-ray crystallography and demonstrated that RC variants exhibit no significant structural alterations relative to wild-type. Ul… Show more

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Cited by 2 publications
(2 citation statements)
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“…Q A is absent in the W­(M252)V X-ray crystal structure (PDB Code 7MHA). Both the W­(M252)V and W­(M252)­V-A­(M260)W RCs show wild-type spectral and primary ET characteristics, the difference from the native RC being loss of Q A activity. All samples had Q B displaced by addition of terbutryn.…”
Section: Methodsmentioning
confidence: 99%
“…Q A is absent in the W­(M252)V X-ray crystal structure (PDB Code 7MHA). Both the W­(M252)V and W­(M252)­V-A­(M260)W RCs show wild-type spectral and primary ET characteristics, the difference from the native RC being loss of Q A activity. All samples had Q B displaced by addition of terbutryn.…”
Section: Methodsmentioning
confidence: 99%
“…Rhodobacter spheroides RCs (M252V mutant which does not assemble with QA) were isolated as previously described. 31 The protein was stored in 10 mM Tris (pH 8.0), containing 0.03% (w/v) LDAO. For reconstitution, a film was prepared in a glass vial by evaporating solvents from 80 µL of 5 mg/mL solution of PG2 in 1:1 CHCl3:MeOH.…”
Section: Expression Purification and Reconstitution Of Photosynthetic...mentioning
confidence: 99%