The Effects of Ionic Strength on Protein Stability: The Cold Shock Protein Family

Dominy, Brian N.; Perl, Dieter; Schmid, Franz X.; Brooks, Charles L.

doi:10.1016/s0022-2836(02)00259-0

Cited by 151 publications

(188 citation statements)

References 44 publications

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“…Monovalent salts such as NaCl modify the ionic strength of solutions, such as the cellular cytoplasm, and can exert stabilizing or destabilizing effects on proteins, with the net effect depending on the nature of specific charge distribution and ionic interactions within the protein (68)(69)(70). Electrolyte concentration also is central to the screening of surface charge-charge interactions that influence protein stability, interaction, and function.…”

Section: Discussionmentioning

confidence: 99%

Ionic imbalance, in addition to molecular crowding, abates cytoskeletal dynamics and vesicle motility during hypertonic stress

Nunes

Roth

Meda

et al. 2015

Proc. Natl. Acad. Sci. U.S.A.

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Cell volume homeostasis is vital for the maintenance of optimal protein density and cellular function. Numerous mammalian cell types are routinely exposed to acute hypertonic challenge and shrink. Molecular crowding modifies biochemical reaction rates and decreases macromolecule diffusion. Cell volume is restored rapidly by ion influx but at the expense of elevated intracellular sodium and chloride levels that persist long after challenge. Although recent studies have highlighted the role of molecular crowding on the effects of hypertonicity, the effects of ionic imbalance on cellular trafficking dynamics in living cells are largely unexplored. By tracking distinct fluorescently labeled endosome/vesicle populations by live-cell imaging, we show that vesicle motility is reduced dramatically in a variety of cell types at the onset of hypertonic challenge. Live-cell imaging of actin and tubulin revealed similar arrested microfilament motility upon challenge. Vesicle motility recovered long after cell volume, a process that required functional regulatory volume increase and was accelerated by a return of extracellular osmolality to isosmotic levels. This delay suggests that, although volume-induced molecular crowding contributes to trafficking defects, it alone cannot explain the observed effects. Using fluorescent indicators and FRET-based probes, we found that intracellular ATP abundance and mitochondrial potential were reduced by hypertonicity and recovered after longer periods of time. Similar to the effects of osmotic challenge, isovolumetric elevation of intracellular chloride concentration by ionophores transiently decreased ATP production by mitochondria and abated microfilament and vesicle motility. These data illustrate how perturbed ionic balance, in addition to molecular crowding, affects membrane trafficking.electrolyte | cell volume | hypertonicity | membrane trafficking | mitochondria

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Section: Discussionmentioning

confidence: 99%

Ionic imbalance, in addition to molecular crowding, abates cytoskeletal dynamics and vesicle motility during hypertonic stress

Nunes

Roth

Meda

et al. 2015

Proc. Natl. Acad. Sci. U.S.A.

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“…Modifying the ionic strength or molarity of a solution can affect the stability of a protein (32,41), depending on its charge distribution (27). Similarly charged residues in a protein, which are screened in solutions containing salt, repel each other in low-salt environments (15,19,31), thereby destabilizing it.…”

Section: Discussionmentioning

confidence: 99%

Soluble Respiratory Syncytial Virus Fusion Protein in the Fully Cleaved, Pretriggered State Is Triggered by Exposure to Low-Molarity Buffer

Chaiwatpongsakorn

Epand

Collins

et al. 2011

J Virol

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The paramyxovirus fusion (F) glycoprotein is anchored in the virion membrane in a metastable, pretriggered form. Once triggered, the F protein undergoes a dramatic conformational extension that inserts its hydrophobic fusion peptide into the target cell membrane, then folds back on itself to bring the membranes together and initiate fusion. Unlike most other paramyxoviruses, the respiratory syncytial virus (RSV) F protein alone is sufficient to mediate membrane fusion and virus infection. To study the triggering mechanism of the RSV F protein, we have generated a soluble F (sF) protein by replacing the transmembrane and cytoplasmic tail domains with a 6His tag. The sF protein is secreted efficiently from 293T cells in a fully cleaved form. It is recognized by neutralizing monoclonal antibodies, appears spherical by electron microscopic analysis, and is not aggregated, all consistent with a native, pretriggered trimer. The sF protein was purified on a Ni ؉2 column and eluted with 50 mM phosphate buffer containing 500 mM NaCl and 250 mM imidazole. Dialysis against 10 mM buffer caused the sF protein to trigger, forming "hat pin"-shaped molecules that aggregated as rosettes, characteristic of the posttriggered form. Further dialysis experiments indicated that the efficiency of triggering correlated well with the reduction of buffer molarity. Reduction of buffer molarity by dilution also resulted in exposure of the fusion peptide, as detected by liposome association, confirming sF protein triggering. Mutation of the furin cleavage site adjacent to the fusion peptide prevented liposome association, further confirming that association is via the fusion peptide.

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“…In the past, many works have been done in this field 1,2 and some physical characters were regarded as the keys to the thermostability of proteins, such as hydrogen bonds and fractional polar surface, 3,4 hydrophobicity, 5,6 electrostatic interactions, [7][8][9][10][11][12] salt bridges, [13][14][15][16] and contact density. 17 Generally it is believed that the electrostatic interaction (include ion pairs and hydrogen bonds) and the hydrophobic interaction (nonpolar residue packing) in the proteins are the most important factors to protein thermostability (as well reviewed in [18][19][20][21][22] ).…”

Section: Introductionmentioning

confidence: 99%

All‐atom contact potential approach to protein thermostability analysis

Chen

Xiao

2006

Biopolymers

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In this paper we use all‐atom potential energy to define and analyze the inter‐residue contacts in mesophilic and thermophilic proteins. Fifteen families of proteins are selected and each family has two representative proteins with greatly different preferred environmental temperatures. We find that both the number and energy of the contacts defined in this way show stronger correlations with the preferred temperatures of proteins than other factors used before. We also find that the charged‐polar and charged‐nonpolar residue contacts not only have larger contact numbers but also have lower single contact energies. Furthermore, the most important is that most of the thermophilic proteins have more charged‐polar and charged‐nonpolar residue contacts than their mesophilic counterparts. This suggests that they may play an important role in the thermostability of proteins, except usual charged–charged and nonpolar–nonpolar residue contacts. Charged residues may exert their profound influence by forming contacts not only with other charged residues but also with polar or nonpolar residues, thus further increasing the strength of contact network and then the thermostability of proteins. © 2006 Wiley Periodicals, Inc. Biopolymers 85: 28–37, 2007.This article was originally published online as an accepted preprint. The “Published Online” date corresponds to the preprint version. You can request a copy of the preprint by emailing the Biopolymers editorial office at biopolymers@wiley.com

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The Effects of Ionic Strength on Protein Stability: The Cold Shock Protein Family

Cited by 151 publications

References 44 publications

Ionic imbalance, in addition to molecular crowding, abates cytoskeletal dynamics and vesicle motility during hypertonic stress

Ionic imbalance, in addition to molecular crowding, abates cytoskeletal dynamics and vesicle motility during hypertonic stress

Soluble Respiratory Syncytial Virus Fusion Protein in the Fully Cleaved, Pretriggered State Is Triggered by Exposure to Low-Molarity Buffer

All‐atom contact potential approach to protein thermostability analysis

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