Abstract
Chloroplasts of a chlorophyll (Chl) b-less barley mutant were solubilized with digitonin and fractionated by polyacrylamide gel electrophoresis with sodium deoxycholate in the running buffer. By this procedure, in contrast to using sodium dodecylsulfate (SDS) for solubilization, a Chl a-protein analogous to the major light-harvesting Chl a-b protein complex from wildtype chloroplasts was recovered. This mutant Chl a-protein comprises about fifty percent of the total Chl a, and is very similar in carotenoid, amino acid, protein and polypeptide composition to the major wildtype antenna Chl a-b protein. The only major differences we have found is its instability in the presence of SDS and sensitivity to protease action. Even with deoxycholate, the mutant Chl a complex often dissociates during electrophoresis into two green bands. The lack of Chl b appears to affect the normal organization of Chl a and protein in such a way as to render the complex more unstable.
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Acker S, Lagoutte B, Picaud A and Duranton J (1982) Protein identification of purified particles isolated from spinach thylakoids by deoxycholate electrophoresis. Photosyn Res 3: 215–225
Apel K and Kloppstech K (1980) The effect of light on the biosynthesis of the light-harvesting chloropbyll a/b protein. Planta 150: 426–430
Bellemare G, Bartlett SG and Chua N-H (1982) Biosynthesis of chlorophyll a/b-binding polypeptides in wild type and the Chlorina f2 mutant of barley. J Biol Chem 257: 7762–7767
Bennett J (1981) Biosynthesis of the light-harvesting chlorophyll a/b protein. Eur J Biochem 118: 61–70
Boardman NK and Highkin HR (1966) Studies on a barley mutant lacking chlorophyll b I. Photochemical activity of isolated chloroplasts. Biochim Biophys Acta 126: 189–199
Boardman NK and Thorne SW (1968) Studies on a barley mutant lacking chlorophyll b II. Fluorescence properties of isolated chloroplasts. Biochim Biophys Acta 153: 448–458
Brown JS (1983) A new evaluation of chlorophyll absorption in photosynthetic membranes. Photosyn Res 4: 375–383
Brown JS and Schoch S (1981) Spectral analysis of chlorophyll protein complexes from higher plant chloroplasts. Biochim Biophys Acta 636: 201–209
Burke JJ, Steinback KE and Arntzen CJ (1979) Analysis of the light-harvesting pigment-protein complex of wild type and a chlorophyll-b-less mutant of barley. Plant Physiol 63: 237–243
Dunkley PR and Anderson JM (1979) The light-harvesting chlorophyll a/b-protein complex from barley thylakoid membranes. Biochim Biophys Acta 545: 174–187
Goodchild DJ, Highkin HR and Boardman NK (1966) The fine structure of chloroplasts in a barley mutant lacking chlorophyll b. Exp Cell Res 43: 684–688
Henriques F and Park RB (1975) Further chemical and morphological characterization of chloroplast membranes from a chlorophyll b-less mutant of Hordeum vulgare. Plant Physiol 55: 763–767
Highkin HR and Frenkel AW (1962) Studies of growth and metabolism of a barley mutant lacking chlorophyll b. Plant Physiol 37: 814–820
Hiyama T and Ke B (1972) Difference spectra and extinction coefficients of P700. Biochim Biophys Acta 267: 160–171
Machold O, Meister A, Sagromsky H, Hoyer-Hansen G and vonWettstein D (1976) Composition of photosynthetic membranes of wild-type barley and chlorophyll b-less mutants. Photosynthetica 11: 200–206.
Mackinney G (1940) Criteria for purity of chlorophyll preparations. J Biol Chem 132: 91–109.
Picaud A, Acker S and Duranton J (1982) A single step separation of Ps 1, Ps 2 and chlorophyll-antenna particles from spinach chloroplasts. Photosyn Res 3: 203–213.
Ryrie IJ (1983) Immunological evidence for apoproteins of the light-harvesting chlorophyll-protein complex in a mutant of barley lacking chlorophyll b. Eur J Biochem 131: 149–155
Ryrie IJ and Fuad N (1982) Membrane adhesion in reconstituted proteoliposomes containing the light-harvesting chlorophyll a/b-protein complex: the role of charged surface groups. Arch Biochem Biophys 214: 475–488
Simpson DJ (1979) Freeze-fracture studies on barley plastid membranes. III. Location of the light-harvesting chlorophyll-protein. Carlsberg Res Commun 44: 305–336
Thornber JP and Highkin HR (1974) Composition of the photosynthetic apparatus of normal barley leaves and a mutant lacking chlorophyll b. Eur J Biochem 41: 109–116
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CIW-DPB No. 917.
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Duranton, J., Brown, J. Evidence for a light-harvesting chlorophyll a-protein complex in a chlorophyll b-less barley mutant. Photosynth Res 11, 141–151 (1987). https://doi.org/10.1007/BF00018272
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DOI: https://doi.org/10.1007/BF00018272