Crystallization of reaction center from Rhodopseudomonas sphaeroides: Preliminary characterization

Research output: Contribution to journalArticle

99 Citations (Scopus)

Abstract

Reaction centers (RCs), integral membrane proteins that mediate the conversion of light into chemical energy, were crystallized by two different vapor diffusion techniques. In one method, small amphipathic molecules (1,2,3-heptanetriol and triethylammonium phosphate) were added to the RCs that had been solubilized in detergent. In the second method, crystallization occurred near the phase boundaries of a two-phase system created by the addition of polyethylene glycol and NaCl to RCs in octyl β-D-glucoside. Several different crystal forms were obtained; two were analyzed by x-ray diffraction. One was monoclinic (space group P2) with β = 105°, and a = 70 Å, b = 105 Å, and c = 85 Å, two RCs per unit cell, and one RC per asymmetric unit; the crystal diffracted to 3.5 Å at 17°C. The other crystal form was orthorhombic (space group C222) with a = 185 Å, b = 170 Å, and c = 105 Å, with eight RCs per unit cell and one RC per asymmetric unit. Reversible light-induced EPR signals of the primary donor (bacteriochlorophyll dimer) showed that the RCs in the crystal were fully active. From the angular dependence of the EPR signal the molecular g anisotropy of the bacteriochlorophyll dimer was deduced to be (64 ± 3) x 10-5. Linear dichroism measurements were performed on the monoclinic crystal. The two bands at 535 and 544 nm assigned to the Q(χ) transitions of the bacteriopheophytins were resolved and preliminary orientations of some of the pigments were obtained.

Original languageEnglish
Pages (from-to)4795-4799
Number of pages5
JournalProceedings of the National Academy of Sciences of the United States of America
Volume81
Issue number15 I
Publication statusPublished - 1984

Fingerprint

Bacteriochlorophylls
Rhodobacter sphaeroides
Crystallization
Light
Anisotropy
Detergents
Membrane Proteins
Phosphates
X-Rays
heptane-1,2,3-triol
octyl-beta-D-glucoside
bacteriopheophytin

ASJC Scopus subject areas

  • General
  • Genetics

Cite this

@article{cd454e894e4142f1b5e6c1886b1a9285,
title = "Crystallization of reaction center from Rhodopseudomonas sphaeroides: Preliminary characterization",
abstract = "Reaction centers (RCs), integral membrane proteins that mediate the conversion of light into chemical energy, were crystallized by two different vapor diffusion techniques. In one method, small amphipathic molecules (1,2,3-heptanetriol and triethylammonium phosphate) were added to the RCs that had been solubilized in detergent. In the second method, crystallization occurred near the phase boundaries of a two-phase system created by the addition of polyethylene glycol and NaCl to RCs in octyl β-D-glucoside. Several different crystal forms were obtained; two were analyzed by x-ray diffraction. One was monoclinic (space group P2) with β = 105°, and a = 70 {\AA}, b = 105 {\AA}, and c = 85 {\AA}, two RCs per unit cell, and one RC per asymmetric unit; the crystal diffracted to 3.5 {\AA} at 17°C. The other crystal form was orthorhombic (space group C222) with a = 185 {\AA}, b = 170 {\AA}, and c = 105 {\AA}, with eight RCs per unit cell and one RC per asymmetric unit. Reversible light-induced EPR signals of the primary donor (bacteriochlorophyll dimer) showed that the RCs in the crystal were fully active. From the angular dependence of the EPR signal the molecular g anisotropy of the bacteriochlorophyll dimer was deduced to be (64 ± 3) x 10-5. Linear dichroism measurements were performed on the monoclinic crystal. The two bands at 535 and 544 nm assigned to the Q(χ) transitions of the bacteriopheophytins were resolved and preliminary orientations of some of the pigments were obtained.",
author = "Allen, {James Paul} and G. Feher",
year = "1984",
language = "English",
volume = "81",
pages = "4795--4799",
journal = "Proceedings of the National Academy of Sciences of the United States of America",
issn = "0027-8424",
number = "15 I",

}

TY - JOUR

T1 - Crystallization of reaction center from Rhodopseudomonas sphaeroides

T2 - Preliminary characterization

AU - Allen, James Paul

AU - Feher, G.

PY - 1984

Y1 - 1984

N2 - Reaction centers (RCs), integral membrane proteins that mediate the conversion of light into chemical energy, were crystallized by two different vapor diffusion techniques. In one method, small amphipathic molecules (1,2,3-heptanetriol and triethylammonium phosphate) were added to the RCs that had been solubilized in detergent. In the second method, crystallization occurred near the phase boundaries of a two-phase system created by the addition of polyethylene glycol and NaCl to RCs in octyl β-D-glucoside. Several different crystal forms were obtained; two were analyzed by x-ray diffraction. One was monoclinic (space group P2) with β = 105°, and a = 70 Å, b = 105 Å, and c = 85 Å, two RCs per unit cell, and one RC per asymmetric unit; the crystal diffracted to 3.5 Å at 17°C. The other crystal form was orthorhombic (space group C222) with a = 185 Å, b = 170 Å, and c = 105 Å, with eight RCs per unit cell and one RC per asymmetric unit. Reversible light-induced EPR signals of the primary donor (bacteriochlorophyll dimer) showed that the RCs in the crystal were fully active. From the angular dependence of the EPR signal the molecular g anisotropy of the bacteriochlorophyll dimer was deduced to be (64 ± 3) x 10-5. Linear dichroism measurements were performed on the monoclinic crystal. The two bands at 535 and 544 nm assigned to the Q(χ) transitions of the bacteriopheophytins were resolved and preliminary orientations of some of the pigments were obtained.

AB - Reaction centers (RCs), integral membrane proteins that mediate the conversion of light into chemical energy, were crystallized by two different vapor diffusion techniques. In one method, small amphipathic molecules (1,2,3-heptanetriol and triethylammonium phosphate) were added to the RCs that had been solubilized in detergent. In the second method, crystallization occurred near the phase boundaries of a two-phase system created by the addition of polyethylene glycol and NaCl to RCs in octyl β-D-glucoside. Several different crystal forms were obtained; two were analyzed by x-ray diffraction. One was monoclinic (space group P2) with β = 105°, and a = 70 Å, b = 105 Å, and c = 85 Å, two RCs per unit cell, and one RC per asymmetric unit; the crystal diffracted to 3.5 Å at 17°C. The other crystal form was orthorhombic (space group C222) with a = 185 Å, b = 170 Å, and c = 105 Å, with eight RCs per unit cell and one RC per asymmetric unit. Reversible light-induced EPR signals of the primary donor (bacteriochlorophyll dimer) showed that the RCs in the crystal were fully active. From the angular dependence of the EPR signal the molecular g anisotropy of the bacteriochlorophyll dimer was deduced to be (64 ± 3) x 10-5. Linear dichroism measurements were performed on the monoclinic crystal. The two bands at 535 and 544 nm assigned to the Q(χ) transitions of the bacteriopheophytins were resolved and preliminary orientations of some of the pigments were obtained.

UR - http://www.scopus.com/inward/record.url?scp=0010948917&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0010948917&partnerID=8YFLogxK

M3 - Article

C2 - 6087348

AN - SCOPUS:0010948917

VL - 81

SP - 4795

EP - 4799

JO - Proceedings of the National Academy of Sciences of the United States of America

JF - Proceedings of the National Academy of Sciences of the United States of America

SN - 0027-8424

IS - 15 I

ER -