Rocket and crossed immunoelectrophoresis of proteins solubilized with sodium dodecyl sulfate

Fg, Plumley; Gw, Schmidt

doi:10.1016/0003-2697(83)90267-1

Cited by 45 publications

(16 citation statements)

References 14 publications

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“…A calibration curve was obtained with Rubisco purified from rice leaves. The relative amount of Cyt f was determined by rocket immunoelectrophoresis after solubilization of the precipitate fraction with lithium dodecyl sulfate, according to the method of Plumley and Schmidt (1983), with a slight modification (Makino et al, 1994a). The specific antibody against Cyt f used was previously prepared (Hidema et al, 1991).…”

Section: Biochemical Assaysmentioning

confidence: 99%

The Effect of Elevated Partial Pressures of CO2 on the Relationship between Photosynthetic Capacity and N Content in Rice Leaves

1997

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The effects of growth CO, levels on the photosynthetic rates; the amounts of ribulose-1,s-bisphosphate carboxylase (Rubisco), chlorophyll (Chl), and cytochrome fi sucrose phosphate synthase activity; and total N content were examined i n young, fully expanded leaves of rice (Oryza sativa 1.). l h e plants were grown hydroponically under two CO, partial pressures of 36 and 1 O0 Pa at three N concentrations. The light-saturated photosynthesis at 36 Pa CO, was lower i n the plants grown in 100 Pa CO, than those grown in 36 Pa CO,. Similarly, the amounts of Rubisco, Chl, and total N were decreased in the leaves of the plants grown in 100 Pa CO,. However, regression analysis showed no differences between the two CO, treatments in the relationship between photosynthesis and total N or i n the relationship between Rubisco and Chl and total N. Although a relative decrease in Rubisco to cytochrome f o r sucrose phosphate synthase was found i n the plants grown in 100 Pa C 0 2 , this was the result of a decrease in total N content by CO, enrichment. The activation state of Rubisco was also unaffected by growth CO, levels. Thus, decreases i n the photosynthetic capacity of the plants grown in 100 Pa CO, could be simply accounted for by a decrease in the absolute amount of leaf N. Leaf photosynthesis is affected by the levels of environmental CO, in which the plants are grown. Short-term (seconds to hours) CO, enrichment stimulates the photosynthetic rate per unit of leaf area. Plant mass is also enhanced during a subsequent long-term exposure (weeks to months) to elevated CO, levels. However, such a longterm CO, enrichment reduces the initial stimulation of photosynthesis and then frequently suppresses photosynthesis (for reviews, see Stitt, 1991; Bowes, 1993;Sage, 1994). These findings indicate that prolonged exposure to elevated CO, leads to changes in biochemical, physiological, or morphological factors, which remove or offset the initial stimulation of photosynthesis. However, the mechanisms of the suppression of photosynthesis by long-term CO, enrichment still remain unclear. Growth under high CO, partial pressures leads to carbohydrate accumulation. Since an apparent correlation between carbohydrate accumulation and the suppression of photosynthesis has often been reported (Sasek et al., 1985;Peet et al., 1986;Yelle et al., 1989;Wong, 1990;Xu et al., 1994), much attention has been paid to the causal relationship(s) between the two phenomena. Stitt (1991) proposed the existence of a feedback mechanism(s) by which the accumulation of carbohydrate indirectly leads to a decrease in the amounts of key components of the photosynthetic apparatus. In fact, many studies of long-term acclimation to elevated CO, partial pressures have shown a decrease in Rubisco (Peet et al., 1986;Sage et al., 1989; RowlandBamford et al., 1991;Xu et al., 1994; Jacob et al., 1995; Rogers et al., 1996). In addition, a decrease in the transcript levels of rbcS and rbcL mRNA was recently found in the plants grown at elevated CO, partial pressu...

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Section: Biochemical Assaysmentioning

confidence: 99%

The Effect of Elevated Partial Pressures of CO2 on the Relationship between Photosynthetic Capacity and N Content in Rice Leaves

1997

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“…Qualitative immunochemical analysis was done by double immunodiffusion as described by Oakley (20). Quantitative analysis was done by rocket immunoelectrophoresis by the procedures of Weeke (38) and Plumley and Schmidt (23). The agarose in Tris-glycine buffer (pH 8.6) was supplemented with 1 mM calcium lactate and 4% polyethylene glycol 6000.…”

Section: Methodsmentioning

confidence: 99%

Purification and properties of a protein linked to the soluble hydrogenase of hydrogen-oxidizing bacteria

Kärst¹,

Suetin²,

Friedrich³

1987

J Bacteriol

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In Alcaligenes eutrophus, the formation of the hydrogenases and of five new peptides is subject to the hydrogenase control system. Of these, the B peptide was purified to homogeneity. This protein (Mr, 37,500) was composed of two identical subunits (Mr,18,800). Antibodies against the B protein were used for its quantification by rocket immunoelectrophoresis. About 4% of the total protein consisted of the B protein; its molar ratio to the NAD-linked hydrogenase was about 4:1. The B protein appeared to be associated with the NAD-linked hydrogenase, as shown by gel filtration analysis with Sephadex G-200. The B protein was not detected in cells that had not expressed the hydrogenase proteins or that lacked the genetic information of the hydrogen-oxidizing character; it was also not detected in Tn5 insertional mutants that were unable to form soluble hydrogenase antigens. Immunochemical analysis of other species and genera than A. eutrophus revealed that only strains able to form a NAD-linked hydrogenase also formed B-protein antigens. The B protein is not required for the catalytic activity of soluble hydrogenase in vitro; its function is at present unknown.

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“…In this gel system Dl migrates faster than D2 (13). For (20). Phosphorylated thylakoid proteins of spinach were resolved by SDS-PAGE with urea and then electrophoresed transversely into an agarose gel containing one of the antisera (x 12-x25 dilution) raised against CP47, CP43, Dl, or D2.…”

Section: And Methodsmentioning

confidence: 99%

Phosphorylation of Photosystem II Components, CP43 Apoprotein, D1, D2, and 10 to 11 Kilodalton Protein in Chloroplast Thylakoids of Higher Plants

Ikeuchi¹,

Fg²,

Inoue³

et al. 1987

Plant Physiol.

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ABSTRACIPhosphorylated thylakoid proteins of spinach (Spinacia okracea L.) and pea (Pisum sativum L.) were solubilized, fractionated by sucrose density gradient centrifugation, and analyzed by gel electrophoresis and crossed immunoelectrophoresis to identify the phosphoproteins. It was found that in addition to intense phosphorylation of light-harvesting chlorophyll complex II, four photosystem II components, CP43 apoprotein, Dl, D2, and a 10 to 11 kilodalton protein, are substantially phosphorylated in the light. Furthermore, the CP43 apoprotein, Dl and D2 can be resolved into two electrophoretic subspecies, only one of which is phosphorylated. This indicates that only a fraction of the PSII polypeptides is phosphorylated. Finally, analysis of detergent procedures suggests that the 10 to 11 kilodalton phosphoprotein is a peripheral component of the Orevolving PSII reaction center complex.In studies of the substrates of thylakoid protein kinases, Steinback et aL (22) identified (7, 14).

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Rocket and crossed immunoelectrophoresis of proteins solubilized with sodium dodecyl sulfate

Cited by 45 publications

References 14 publications

The Effect of Elevated Partial Pressures of CO2 on the Relationship between Photosynthetic Capacity and N Content in Rice Leaves

The Effect of Elevated Partial Pressures of CO2 on the Relationship between Photosynthetic Capacity and N Content in Rice Leaves

Purification and properties of a protein linked to the soluble hydrogenase of hydrogen-oxidizing bacteria

Phosphorylation of Photosystem II Components, CP43 Apoprotein, D1, D2, and 10 to 11 Kilodalton Protein in Chloroplast Thylakoids of Higher Plants

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