### Abstract

The stable tyrosine radical Y_{D}^{•} (tyrosine 160 in the D2 polypeptide) in photosystem II (PSII) exhibits nonexponential electron spin-lattice relaxation transients at low temperature. As previously reported, the tetranuclear Mn complex in PSII significantly enhances the spin-lattice relaxation of Y_{D}^{•}. However, in Mn-depleted PSII membranes, the spin-lattice relaxation transients of Y_{D}^{•} are also nonexponential, and progressive power saturation (P_{1/2}) experiments show that it does not behave like an isolated tyrosine radical. A model is developed to treat the interaction of two paramagnets in a rigid lattice at a fixed distance apart but with a random orientation in a magnetic field. This model describes the spin-lattice relaxation of a radical in proximity to another paramagnetic site in terms of three relaxation rate constants: the "intrinsic" relaxation rate, the relaxation rate due to scalar exchange, and the relaxation rate due to dipole-dipole interactions. The intrinsic and the scalar exchange relaxation rates are isotropic and together contribute a single rate constant to the spin-lattice relaxation transients. However, the dipolar relaxation rate is orientation dependent. Each orientation contributes a different dipolar relaxation rate constant to the net spin-lattice relaxation rate constant. The result is a superposition of single-exponential recoveries, each with a different net rate constant, causing the observed saturation-recovery transients to be non-(single)-exponential. Saturation-recovery relaxation transients of Y_{D}^{•} are compared with those of a model tyrosine radical, generated by UV photolysis of L-tyrosine in a borate glass. From this comparison, we conclude that scalar exchange does not make a significant contribution to the spin-lattice relaxation of Y_{D}^{•} in Mn-depleted PSII. We account for the nonexponential relaxation transients obtained from Y_{D}^{•} in Mn-depleted PSII membranes in terms of dipolar-induced relaxation enhancement from the non-heme Fe(II). From simulations of the spin-lattice relaxation transients, we obtain the magnitude of the magnetic dipolar interaction between Y_{D}^{•} and the non-heme Fe(II), which can be used to calculate the distance between them. Using data on the non-heme Fe(II) in the reaction center of Rhodobacter sphaeroides to model the non-heme Fe(II) in PSII, we calculate a Y_{D}^{•}-Fe(II) distance of ≥38 Å in PSII. This agrees well with the distance predicted from the structure of the bacterial reaction center.

Original language | English |
---|---|

Pages (from-to) | 532-541 |

Number of pages | 10 |

Journal | Biochemistry |

Volume | 31 |

Issue number | 2 |

Publication status | Published - 1992 |

### Fingerprint

### ASJC Scopus subject areas

- Biochemistry

### Cite this

*Biochemistry*,

*31*(2), 532-541.

**Using saturation-recovery EPR to measure distances in proteins : Applications to photosystem II.** / Hirsh, Donald J.; Beck, Warren F.; Innes, Jennifer B.; Brudvig, Gary W.

Research output: Contribution to journal › Article

*Biochemistry*, vol. 31, no. 2, pp. 532-541.

}

TY - JOUR

T1 - Using saturation-recovery EPR to measure distances in proteins

T2 - Applications to photosystem II

AU - Hirsh, Donald J.

AU - Beck, Warren F.

AU - Innes, Jennifer B.

AU - Brudvig, Gary W

PY - 1992

Y1 - 1992

N2 - The stable tyrosine radical YD• (tyrosine 160 in the D2 polypeptide) in photosystem II (PSII) exhibits nonexponential electron spin-lattice relaxation transients at low temperature. As previously reported, the tetranuclear Mn complex in PSII significantly enhances the spin-lattice relaxation of YD•. However, in Mn-depleted PSII membranes, the spin-lattice relaxation transients of YD• are also nonexponential, and progressive power saturation (P1/2) experiments show that it does not behave like an isolated tyrosine radical. A model is developed to treat the interaction of two paramagnets in a rigid lattice at a fixed distance apart but with a random orientation in a magnetic field. This model describes the spin-lattice relaxation of a radical in proximity to another paramagnetic site in terms of three relaxation rate constants: the "intrinsic" relaxation rate, the relaxation rate due to scalar exchange, and the relaxation rate due to dipole-dipole interactions. The intrinsic and the scalar exchange relaxation rates are isotropic and together contribute a single rate constant to the spin-lattice relaxation transients. However, the dipolar relaxation rate is orientation dependent. Each orientation contributes a different dipolar relaxation rate constant to the net spin-lattice relaxation rate constant. The result is a superposition of single-exponential recoveries, each with a different net rate constant, causing the observed saturation-recovery transients to be non-(single)-exponential. Saturation-recovery relaxation transients of YD• are compared with those of a model tyrosine radical, generated by UV photolysis of L-tyrosine in a borate glass. From this comparison, we conclude that scalar exchange does not make a significant contribution to the spin-lattice relaxation of YD• in Mn-depleted PSII. We account for the nonexponential relaxation transients obtained from YD• in Mn-depleted PSII membranes in terms of dipolar-induced relaxation enhancement from the non-heme Fe(II). From simulations of the spin-lattice relaxation transients, we obtain the magnitude of the magnetic dipolar interaction between YD• and the non-heme Fe(II), which can be used to calculate the distance between them. Using data on the non-heme Fe(II) in the reaction center of Rhodobacter sphaeroides to model the non-heme Fe(II) in PSII, we calculate a YD•-Fe(II) distance of ≥38 Å in PSII. This agrees well with the distance predicted from the structure of the bacterial reaction center.

AB - The stable tyrosine radical YD• (tyrosine 160 in the D2 polypeptide) in photosystem II (PSII) exhibits nonexponential electron spin-lattice relaxation transients at low temperature. As previously reported, the tetranuclear Mn complex in PSII significantly enhances the spin-lattice relaxation of YD•. However, in Mn-depleted PSII membranes, the spin-lattice relaxation transients of YD• are also nonexponential, and progressive power saturation (P1/2) experiments show that it does not behave like an isolated tyrosine radical. A model is developed to treat the interaction of two paramagnets in a rigid lattice at a fixed distance apart but with a random orientation in a magnetic field. This model describes the spin-lattice relaxation of a radical in proximity to another paramagnetic site in terms of three relaxation rate constants: the "intrinsic" relaxation rate, the relaxation rate due to scalar exchange, and the relaxation rate due to dipole-dipole interactions. The intrinsic and the scalar exchange relaxation rates are isotropic and together contribute a single rate constant to the spin-lattice relaxation transients. However, the dipolar relaxation rate is orientation dependent. Each orientation contributes a different dipolar relaxation rate constant to the net spin-lattice relaxation rate constant. The result is a superposition of single-exponential recoveries, each with a different net rate constant, causing the observed saturation-recovery transients to be non-(single)-exponential. Saturation-recovery relaxation transients of YD• are compared with those of a model tyrosine radical, generated by UV photolysis of L-tyrosine in a borate glass. From this comparison, we conclude that scalar exchange does not make a significant contribution to the spin-lattice relaxation of YD• in Mn-depleted PSII. We account for the nonexponential relaxation transients obtained from YD• in Mn-depleted PSII membranes in terms of dipolar-induced relaxation enhancement from the non-heme Fe(II). From simulations of the spin-lattice relaxation transients, we obtain the magnitude of the magnetic dipolar interaction between YD• and the non-heme Fe(II), which can be used to calculate the distance between them. Using data on the non-heme Fe(II) in the reaction center of Rhodobacter sphaeroides to model the non-heme Fe(II) in PSII, we calculate a YD•-Fe(II) distance of ≥38 Å in PSII. This agrees well with the distance predicted from the structure of the bacterial reaction center.

UR - http://www.scopus.com/inward/record.url?scp=0026580066&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0026580066&partnerID=8YFLogxK

M3 - Article

C2 - 1310040

AN - SCOPUS:0026580066

VL - 31

SP - 532

EP - 541

JO - Biochemistry

JF - Biochemistry

SN - 0006-2960

IS - 2

ER -