Release of Two Peripheral Proteins from Chloroplast Thylakoid Membranes in the Presence of a Hofmeister Series of Chaotropic Anions

Hincha, Dirk K.

doi:10.1006/abbi.1998.0866

Cited by 17 publications

(11 citation statements)

References 38 publications

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“…Alternatively, it represents a loosely attached peripheral PM protein, since detergents at low (<CMC) concentrations can strip membranes of loosely attached components. It is known that high salt concentrations also may cause release of peripheral proteins from membranes (Hatefi and Hanstein 1974, Hincha 1998), but treatment of inside‐out PM vesicles with NaCl did not appear to result in any further release of protein (Fig. 6).…”

Section: Resultsmentioning

confidence: 91%

Protein complexes of the plant plasma membrane resolved by Blue Native PAGE

Kjell

Rasmusson

Larsson

et al. 2004

Physiologia Plantarum

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With the characterization of the total genomes of Arabidopsis thaliana and Oryza sativa, several putative plasma membrane components have been identified. However, a lack of knowledge at the protein level, especially for hydrophobic proteins, have hampered analyses of physiological changes. To address whether protein complexes may be present in the native membrane, we subjected plasma membranes isolated from Spinacia oleracea leaves to blue‐native polyacrylamide gel electrophoresis (BN‐PAGE). BN‐PAGE is well established in the separation of functional membrane protein complexes from mitochondria and chloroplasts, but a resolved protein complex pattern from PM of eukaryotic cells has previously not been reported. Using this method, protein complexes from Spinacia oleracea PM could be efficiently solubilized and separated, including the highly hydrophobic aquaporin (apparent molecular mass 230 kDa), a putative tetramer of H+‐ATPase, and several less abundant complexes with apparent masses around or above 750 kDa. After denaturation and separation of the complexes into their subunits in a second dimension (SDS‐PAGE), several of the complexes were identified as hydrophobic membrane proteins. Large amounts of protein (up to 1 mg) can be resolved in each lane, which suggests that the method could be used to study also low‐abundance protein complexes, e.g. under different physiological conditions.

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Section: Resultsmentioning

confidence: 91%

Protein complexes of the plant plasma membrane resolved by Blue Native PAGE

Kjell

Rasmusson

Larsson

et al. 2004

Physiologia Plantarum

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“…The protection provided by sorbitol was found to be much smaller than that provided by an equal amount of the natural compatible solute glycerol (Shen et al, 1999). The direct protection of soluble and peripherally bound membrane proteins at high osmotic stress is described as a key factor for the efficiency of an osmotic protectant, which has been demonstrated for GG (Santoro et al, 1992;Hincha, 1998;Borges et al, 2002).…”

Section: Discussionmentioning

confidence: 98%

Osmotic stress in Synechocystis sp. PCC 6803: low tolerance towards nonionic osmotic stress results from lacking activation of glucosylglycerol accumulation

et al. 2006

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In order to compare the molecular principles of the acclimatization of bacterial cells to salt and nonionic osmotic stress, the moderately halotolerant cyanobacterium Synechocystis sp. PCC 6803 was challenged by salt (NaCl), and the osmolytes sorbitol and maltose. The physiological response towards each of the three compounds was found to be different. After salt addition, the cell volume remained unchanged, and the accumulation of the osmoprotective compound glucosylglycerol (GG) was observed after activation of the key enzyme GgpS at the biochemical and gene (ggpS) expression level. Sorbitol addition had only minor effects on the cell volume. In spite of the fact that the ggpS expression was increased, the GgpS enzyme was not activated, resulting in the absence of GG accumulation. In contrast the cells accumulated sorbitol, which served as a compatible solute and assured a certain osmotic resistance. In comparison to NaCl and sorbitol, the addition of maltose caused a strong decrease in cell volume indicating water efflux. However, no osmolyte accumulation was observed, resulting in an osmosensitive phenotype. Consequently, a successful response of Synechocystis cells to an osmotic challenge is indicative of the de novo synthesis of GG upon salt-dependent activation of the GgpS enzyme or the uptake of external solutes.

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“…This implies that membrane stability may not be the limiting factor for these cells under salt stress. In addition to purely osmotic effects, solutes such as Bet, but also Suc and Tre, have been shown to be extremely efficient in stabilizing soluble proteins [16,53] and peripherally bound membrane proteins [54][55][56] under stress conditions and this may also contribute to cellular salt stress tolerance. Cyanobacteria showing high tolerance to desiccation, on the other hand, accumulate Suc and Tre [57,58], which perfectly matches their membrane-protective properties.…”

Section: Discussionmentioning

confidence: 99%

Stabilization of model membranes during drying by compatible solutes involved in the stress tolerance of plants and microorganisms

Hincha

Hagemann

2004

Biochemical Journal

174

109

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Many organisms accumulate compatible solutes under environmental stress conditions. Cyanobacteria accumulate compatible solutes in response to increased external salinity, with tolerance increasing from Suc (sucrose) or trehalose to 2-O-(alpha-D-glucopyranosyl)-glycerol and glycinebetaine accumulating species. It is not clear how these different solutes influence salt tolerance. One possible explanation may be a differential ability of these solutes to stabilize membranes under stress conditions. We therefore performed drying experiments with liposomes in the presence of compatible solutes. Suc, trehalose and sorbitol protected liposomes from leakage of a soluble marker and from membrane fusion during drying and rehydration. 2-O-(alpha-D-glucopyranosyl)-glycerol was less effective and glycinebetaine showed hardly any effect. In combination with Suc, the latter two solutes showed improved protection. Lipid-phase transitions are known to contribute to solute leakage from liposomes. We determined phase transitions in dry membranes in the absence or presence of the solutes, using Fourier-transform infrared spectroscopy. The ability of the solutes to decrease the phase transition temperature corresponded closely to their ability to protect the liposomes against solute leakage. All solutes interacted with the phosphate in the lipid headgroups. The magnitude of the shift in the asymmetric P=O stretching vibration correlated closely with the lipid-phase transition temperature. This indicates that the degree of membrane protection afforded by the solutes is mainly determined by their ability to interact with the membrane lipids. However, this is not a determinant of cellular protection against salt stress, as the solutes show a reverse order when ranked with regard to protection against these stresses.

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Release of Two Peripheral Proteins from Chloroplast Thylakoid Membranes in the Presence of a Hofmeister Series of Chaotropic Anions

Cited by 17 publications

References 38 publications

Protein complexes of the plant plasma membrane resolved by Blue Native PAGE

Protein complexes of the plant plasma membrane resolved by Blue Native PAGE

Osmotic stress in Synechocystis sp. PCC 6803: low tolerance towards nonionic osmotic stress results from lacking activation of glucosylglycerol accumulation

Stabilization of model membranes during drying by compatible solutes involved in the stress tolerance of plants and microorganisms

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