Carbohydrate Protection of Enzyme Structure and Function against Guanidinium Chloride Treatment Depends on the Nature of Carbohydrate and Enzyme

Sola‐Penna, Mauro; Ferreira‐Pereira, Antônio; Lemos, Adriana dos Passos; Meyer-Ferwandes, José Roberto

doi:10.1111/j.1432-1033.1997.00024.x

Cited by 68 publications

(48 citation statements)

References 42 publications

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“…Materials-RNase A (bovine pancreatic), lysozyme (hen egg white, HEW), ␣-CTgen (bovine pancreatic), cyt c (horse heart, type IV), TrpInh (HEW), trehalose, and 2Ј,3Ј-cCMP (NH 4 ϩ salt) were all from Sigma Chemical Co., St. Louis, MO. All the proteins were dialyzed against distilled de-ionized water and lyophilized followed by drying over P 2 O 5 .…”

Section: Methodsmentioning

confidence: 99%

“…Sugars have been known to protect proteins against loss of activity (1, 2), chemical (3,4), and thermal denaturation (5)(6)(7)(8)(9). Among several sugars, ␣,␣-trehalose (␣-D-glucopyranosyl(131)-␣-D-glucopyranoside) has been known to be a superior stabilizer in providing protection to biological materials against dehydration and desiccation (10,11).…”

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confidence: 99%

“…Different proteins are expected to interact with cosolvent molecules in varied ways depending on their physicochemical properties. In general, trehalose has been observed to provide protection to different proteins to various extents and the efficacy of protection depends on the nature of the protein (2,4). Despite the availability of such data, the exact role of proteins and their physico-chemical properties in trehalose-mediated stability is still not clear.…”

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Why Is Trehalose an Exceptional Protein Stabilizer?

Kaushik¹,

Bhat

2003

Journal of Biological Chemistry

555

246

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Trehalose, a naturally occurring osmolyte, is known to be an exceptional stabilizer of proteins and helps retain the activity of enzymes in solution as well as in the freeze-dried state. To understand the mechanism of action of trehalose in detail, we have conducted a thorough investigation of its effect on the thermal stability in aqueous solutions of five well characterized proteins differing in their various physico-chemical properties. Among them, RNase A has been used as a model enzyme to investigate the effect of trehalose on the retention of enzymatic activity upon incubation at high temperatures. 2 M trehalose was observed to raise the transition temperature, T m of RNase A by as much as 18°C and Gibbs free energy by 4.8 kcal mol ؊1 at pH 2.5. There is a decrease in the heat capacity of protein denaturation (⌬C p ) in trehalose solutions for all the studied proteins. An increase in the ⌬G and a decrease in the ⌬C p values for all the proteins points toward a general mechanism of stabilization due to the elevation and broadening of the stability curve (⌬G versus T). A direct correlation of the surface tension of trehalose solutions and the thermal stability of various proteins has been observed. Wyman linkage analysis indicates that at 1.5 M concentration 4 -7 molecules of trehalose are excluded from the vicinity of protein molecules upon denaturation. We further show that an increase in the stability of proteins in the presence of trehalose depends upon the length of the polypeptide chain. The pH dependence data suggest that even though the charge status of a protein contributes significantly, trehalose can be expected to work as a universal stabilizer of protein conformation due to its exceptional effect on the structure and properties of solvent water compared with other sugars and polyols.Sugars have been known to protect proteins against loss of activity (1, 2), chemical (3, 4), and thermal denaturation (5-9). Among several sugars, ␣,␣-trehalose (␣-D-glucopyranosyl(131)-␣-D-glucopyranoside) has been known to be a superior stabilizer in providing protection to biological materials against dehydration and desiccation (10, 11). It is a compatible osmolyte that gets accumulated in organisms under stress conditions (12, 13). Because of this unique property, tremendous interest has been generated in understanding the molecular basis of stress management through induction of trehalose biosynthesis (13, 14).Trehalose has also been found to be very effective in the stabilization of labile proteins during lyophilization (15, 16) and exposure to high temperatures in solution (2,8,9). Sugars in general protect proteins against dehydration by hydrogen bonding to the dried protein by serving as water substitute (15, 17). Several studies carried out by Timasheff and coworkers (9,18) show that sugars and polyols stabilize the folded structure of proteins in solution as a result of greater preferential hydration of the unfolded state compared with the native state. The mechanism is fundamentally different from stabil...

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Section: Methodsmentioning

confidence: 99%

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confidence: 99%

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Why Is Trehalose an Exceptional Protein Stabilizer?

Kaushik¹,

Bhat

2003

Journal of Biological Chemistry

555

246

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“…It stabilizes the partially unfolded state of the protein (rather than stabilizing the native structure). 83 Trehalose is thought to replace the water shell around proteins/membranes and to preserve the three-dimensional structure of the biomolecule. 75 There is a huge body of evidence in support of this viewpoint as well, especially when proteins are freeze-dried for preservation.…”

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Effect of trehalose on protein structure

Jain¹,

Roy²

2008

Protein Science

733

576

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Trehalose is a ubiquitous molecule that occurs in lower and higher life forms but not in mammals. Till about 40 years ago, trehalose was visualized as a storage molecule, aiding the release of glucose for carrying out cellular functions. This perception has now changed dramatically. The role of trehalose has expanded, and this molecule has now been implicated in a variety of situations. Trehalose is synthesized as a stress-responsive factor when cells are exposed to environmental stresses like heat, cold, oxidation, desiccation, and so forth. When unicellular organisms are exposed to stress, they adapt by synthesizing huge amounts of trehalose, which helps them in retaining cellular integrity. This is thought to occur by prevention of denaturation of proteins by trehalose, which would otherwise degrade under stress. This explanation may be rational, since recently, trehalose has been shown to slow down the rate of polyglutamine-mediated protein aggregation and the resultant pathogenesis by stabilizing an aggregation-prone model protein. In recent years, trehalose has also proved useful in the cryopreservation of sperm and stem cells and in the development of a highly reliable organ preservation solution. This review aims to highlight the changing perception of the role of trehalose over the last 10 years and to propose common mechanisms that may be involved in all the myriad ways in which trehalose stabilizes protein structures. These will take into account the structure of trehalose molecule and its interactions with its environment, and the explanations will focus on the role of trehalose in preventing protein denaturation.

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“…On the other hand, the non-stabilizer compounds would make a preferen tial interaction with the protein and it could mobi lize a certain amount of water molecules of the bulk water, increasing the solvation layer. This phenom enon contributes to decrease the forces that stabi lize the tertiary and quaternary structures of the protein, making it more sensitive to changes in the medium (Timasheff, 1993;Sola-Penna et al, 1997;Lopes et al, 1999).…”

Section: Introductionmentioning

confidence: 99%

Carbohydrates Protect Mitochondrial F₀F₁-ATPase Complex against Thermal Inactivation

Saad-Nehme

Silva

Meyer‐Fernandes

2000

Zeitschrift Für Naturforschung C

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Carbohydrates, Thermal Stabilization, Mitochondrial F 0F r ATPaseOrganisms and cellular systems are required to adapt to stress conditions like high tem per ature, often responding by accumulating organic solutes, such as sugars. This accumulation is associated with the effectiveness of these osmolytes in minimizing protein denaturation and membrane damage under stress conditions. In this work, we have studied the effect of sugars on the protection against thermal inactivation of mitochondrial FoFj-ATPase complex, in preparations of submitochondrial particles containing or depleted of inhibitor protein. We observed that after 15 min of pre-incubation at 70 °C of latent MgATP-submitochondrial particles (with inhibitor protein) in the presence of 1.5 m of sucrose or trehalose, or 3.0 m of glucose or fructose, about 80% of enzyme activity remained active. In the same conditions, but in the absence of sugars, the activity of the particles was completely abolished. Submitochondrial particles depleted of the inhibitor protein (A S-particles) were almost completely inactivated after 3 min of pre-incubation at 70 °C in the absence of sugars and more than 60% of the enzyme activity remained active when these particles were pre-incubated in the presence of sugars. In such condition, the enzyme acquires a more com pact and heat-stable conformation. Sugars, as well as the inhibitor protein, inhibit reversibly F 0F r ATPase com plex activity and protect this enzyme against inactivation by high temperature. Interestingly, the protection, promoted by sugars, of particles containing inhibitor protein is higher than of particles depleted of inhibitor protein, suggesting a synergism between sugar and inhibi tor protein.

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Carbohydrate Protection of Enzyme Structure and Function against Guanidinium Chloride Treatment Depends on the Nature of Carbohydrate and Enzyme

Cited by 68 publications

References 42 publications

Why Is Trehalose an Exceptional Protein Stabilizer?

Why Is Trehalose an Exceptional Protein Stabilizer?

Effect of trehalose on protein structure

Carbohydrates Protect Mitochondrial F₀F₁-ATPase Complex against Thermal Inactivation

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Carbohydrate Protection of Enzyme Structure and Function against Guanidinium Chloride Treatment Depends on the Nature of Carbohydrate and Enzyme

Cited by 68 publications

References 42 publications

Why Is Trehalose an Exceptional Protein Stabilizer?

Why Is Trehalose an Exceptional Protein Stabilizer?

Effect of trehalose on protein structure

Carbohydrates Protect Mitochondrial F0F1-ATPase Complex against Thermal Inactivation

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Carbohydrates Protect Mitochondrial F₀F₁-ATPase Complex against Thermal Inactivation