Protein secondary structure of the isolated photosystem II reaction center and conformational changes studied by Fourier transform infrared spectroscopy.
The secondary structure of the photosystem II (PSII) reaction center isolated from pea chloroplasts has been characterized by Fourier transform infrared (FTIR) spectroscopy. Spectra were recorded in aqueous buffers containing H2O or D2O; the detergent present for most measurements was dodecyl maltoside. The broad amide I and amide II bands were analyzed by using second-derivative and deconvolution procedures. Absorption bands were assigned to the presence of alpha-helices, beta-sheets, turns, or random structure. Quantitative analysis revealed that this complex contained a high proportion of alpha-helices (67%) and some antiparallel beta-sheets (9%) and turns (11%). An irreversible decrease in the intensity of the band associated with the alpha-helices occurs upon exposure of the isolated PSII reaction center to bright illumination. This loss of alpha-helical content gave rise to an increase in other secondary structures, particularly beta-sheets. After similar pretreatment with light, sodium dodecyl sulfate polyacrylamide gel electrophoresis reveals lower mobility and solubility of constituent D1 and D2 polypeptides of the PSII reaction center. Some degradation of these polypeptides also occurs. In contrast, there is no change in the mobility of the two subunits of cytochrome b559. In the absence of illumination, the PSII reaction center exchanged into dodecyl maltoside shows good thermal stability as compared with samples in Triton X-100. Only at a temperature of about 60 degrees C do spectral changes take place that are indicative of denaturation.
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W Z
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Wolfson Laboratories, Department of Biochemistry, Imperial College of Science, Technology & Medicine, London, U.K.
Newell
W R
WR
Haris
P I
PI
Chapman
D
D
Barber
J
J
D002021:Buffers; D003573:Cytochrome b Group / Q000737:chemistry; D003902:Detergents / Q000494:pharmacology; D003903:Deuterium; D017666:Deuterium Oxide; D007887:Fabaceae / Q000032:analysis; D005960:Glucosides / Q000494:pharmacology; D017830:Octoxynol; D045322:Photosynthetic Reaction Center Complex Proteins / Q000737:chemistry*; D045332:Photosystem II Protein Complex; D010946:Plants, Medicinal; D011092:Polyethylene Glycols / Q000494:pharmacology; D011487:Protein Conformation / Q000187:drug effects; D011489:Protein Denaturation; D012995:Solubility; D013055:Spectrophotometry, Infrared; D014867:Water
D002021:Buffers; D003573:Cytochrome b Group; D003902:Detergents; D005960:Glucosides; D045322:Photosynthetic Reaction Center Complex Proteins; D045332:Photosystem II Protein Complex; D014867:Water; D011092:Polyethylene Glycols; C040358:dodecyl maltoside; D017830:Octoxynol; C019313:cytochrome b559; D003903:Deuterium; D017666:Deuterium Oxide