The role of trehalose synthesis for the acquisition of thermotolerance in yeast. II. Physiological concentrations of trehalose increase the thermal stability of proteins in vitro.
In baker's yeast (Saccharomyces cerevisiae), accumulation of the non-reducing disaccharide, trehalose, is triggered by stimuli that activate the heat-shock response. Previously, trehalose levels have been shown to be closely correlated with thermotolerance, suggesting a protective function of this substance. Genetic evidence in support of this view is presented in an accompanying paper [De Virgilio, C., Hottiger, T., Dominguez, J., Boller, T. & Wiemken, A. (1993) Eur. J. Biochem. 219, 179-186]. In this study, we have examined the effect of trehalose on the thermal stability of proteins, a parameter thought to be a major determinant of thermotolerance. Physiological concentrations of trehalose (up to 0.5 M) were found to efficiently protect enzymes of yeast (glucose-6P-dehydrogenase, phosphoglucose-isomerase) as well as enzymes of non-yeast origin (bovine glutamic dehydrogenase, EcoRI) against heat inactivation in vitro. Trehalose also reduced the heat-induced formation of protein aggregates. The disaccharide proved to be a compatible solute, as even at very high concentrations (up to 1 M) it did not significantly interfere with the activity of test enzymes. Trehalose was at least as good or better a protein stabilizer than any of a number of other compatible solutes (including sugars, polyalcohols and amino acids), while the structurally related trehalose-6P was devoid of any protective effect. Thermoprotection of enzymes by trehalose was evident even in solutions containing high concentrations of yeast protein or substrate. The data indicate that trehalose accumulation may increase the thermotolerance of yeast by enhancing protein stability in intact cells.
Hottiger
T
T
Botanisches Institut, Universität Basel, Switzerland.
De Virgilio
C
C
Hall
M N
MN
Boller
T
T
Wiemken
A
A
D000064:Acclimatization / Q000502:physiology; D000818:Animals; D002417:Cattle; D015246:Deoxyribonuclease EcoRI / Q000737:chemistry / Q000378:metabolism; D004795:Enzyme Stability; D005656:Fungal Proteins / Q000378:metabolism*; D005956:Glucose-6-Phosphate Isomerase / Q000737:chemistry / Q000378:metabolism*; D005954:Glucosephosphate Dehydrogenase / Q000737:chemistry / Q000378:metabolism*; D005969:Glutamate Dehydrogenase / Q000737:chemistry / Q000378:metabolism; D006358:Hot Temperature; D007700:Kinetics; D012441:Saccharomyces cerevisiae / Q000254:growth & development / Q000378:metabolism / Q000502:physiology*; D013816:Thermodynamics; D014199:Trehalose / Q000378:metabolism / Q000494:pharmacology / Q000502:physiology*
D005656:Fungal Proteins; D014199:Trehalose; D005954:Glucosephosphate Dehydrogenase; D005969:Glutamate Dehydrogenase; D015246:Deoxyribonuclease EcoRI; D005956:Glucose-6-Phosphate Isomerase