DOI: 10.1111/j.1432-1033.1989.tb14870.x. Characterization of trehalose‐6‐phosphate synthase and trehalose‐6‐phosphate phosphatase of <i>Saccharomyces cerevisiae</i>

Characterization of trehalose‐6‐phosphate synthase and trehalose‐6‐phosphate phosphatase of Saccharomyces cerevisiae

10.1111/j.1432-1033.1989.tb14870.x

Crossref journal-article

Wiley

European Journal of Biochemistry (311)

Abstract

The properties of yeast trehalose‐6‐phosphate synthase were reinvestigated in relation with the recent claim made by Panek et al. [Panek, A. C., de Araujo, P. S., Moura‐Neto, V. and Panek, A. D. (1987) Curr. Genet. 11, 459–465] that the enzyme would be stimulated by ATP and partially inactivated by cAMP‐dependent protein kinase. Trehalose‐6‐phosphate synthase activity was measured by the sum of [14C]trehalose 6‐phosphate and [14C]trehalose formed from UDP‐[14C]glucose and glucose 6‐phosphate. The activity measured in an extract of Saccharomyces cerevisiae was not affected by any treatment of the cells, such as incubation in the presence of glucose or of dinitrophenol, which are known to greatly increase the intracellular concentration of cAMP, nor by preincubation of the extract in the presence of ATP‐Mg, cAMP and bovine heart cAMP‐dependent protein kinase. The activity was also not significantly different in several mutants affected in the cAMP system. The kinetic properties of the partially purified enzyme were investigated; no effect of ATP could be detected but Pi acted as a potent noncompetitive inhibitor (Ki= 2 mM). The activity of trehalose‐6‐phosphate phosphatae was measured by the amount of [14C]trehalose formed from [14C]trehalose 6‐phosphate. The enzyme could be separated from other phosphatases and appeared to be highly specific for trehalose 6‐phosphate. It was Mg dependent and its kinetics for trehalose 6‐phosphate was hyperbolic. Studies performed with intact cells, crude extracts or the purified enzyme did not reveal any cAMP‐dependent change in its activity.Remarkably, trehalose‐6‐phosphate synthase and trehalose‐6‐phosphate phosphatase copurified in the course of different chromatographic procedures, suggesting that they are part of a single bifunctional protein. A 50‐fold purification of the two enzymes could be achieved with a yeild of only 2% by chromatography on Mono S followed by gel filtration on Superose 6B.

Bibliography

VANDERCAMMEN, A., FRANÃOIS, J., & HERS, H. (1989). Characterization of trehaloseâ6âphosphate synthase and trehaloseâ6âphosphate phosphatase of Saccharomyces cerevisiae. European Journal of Biochemistry, 182(3), 613â620. Portico.

Authors 3

Annick VANDERCAMMEN (first)
Jean FRANÇOIS (additional)
Henri‐Géry HERS (additional)

References 23 Referenced 107

10.1016/S0065-2318(08)60266-8
10.1016/S0021-9258(19)77303-7
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10.1111/j.1432-1033.1967.tb00104.x
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10.1111/j.1432-1033.1988.tb13830.x
10.1016/0003-2697(76)90527-3 / Anal. Biochem. by Brandford M. A. (1976)
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10.1128/jb.143.3.1384-1394.1980 / J. Bacteriol. by Lillie S. H. (1981)
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10.1002/j.1460-2075.1985.tb03981.x
{'key': 'e_1_2_3_15_2', 'first-page': '239', 'volume-title': 'The enzymes', 'author': 'Bloxham D. P.', 'year': '1973'} / The enzymes by Bloxham D. P. (1973)
10.1016/S0300-9084(73)80393-1
{'key': 'e_1_2_3_17_2', 'first-page': '560', 'volume': '174', 'author': 'François J.', 'year': '1988', 'journal-title': 'Eur. J. Biochem.'} / Eur. J. Biochem. by François J. (1988)
10.1016/S0021-9258(17)39970-2
10.1016/0003-9861(57)90215-1
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10.1128/jb.107.1.217-222.1971
10.1111/j.1432-1033.1986.tb09877.x
10.1016/S0021-9258(18)64469-2

Dates

Type	When
Created	20 years, 5 months ago (March 3, 2005, 10:30 p.m.)
Deposited	1 year, 9 months ago (Nov. 23, 2023, 5:54 p.m.)
Indexed	1 year ago (Aug. 9, 2024, 12:32 p.m.)
Issued	36 years, 1 month ago (July 1, 1989)
Published	36 years, 1 month ago (July 1, 1989)
Published Online	20 years, 5 months ago (March 3, 2005)
Published Print	36 years, 1 month ago (July 1, 1989)

Funders 0

None

BibTeX

@article{VANDERCAMMEN_1989, title={Characterization of trehalose‐6‐phosphate synthase and trehalose‐6‐phosphate phosphatase of Saccharomyces cerevisiae}, volume={182}, ISSN={1432-1033}, url={http://dx.doi.org/10.1111/j.1432-1033.1989.tb14870.x}, DOI={10.1111/j.1432-1033.1989.tb14870.x}, number={3}, journal={European Journal of Biochemistry}, publisher={Wiley}, author={VANDERCAMMEN, Annick and FRANÇOIS, Jean and HERS, Henri‐Géry}, year={1989}, month=jul, pages={613–620} }

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