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Volume 137, Issue 2

Trehalose-6-phosphate synthase/phosphatase complex from bakers’ yeast: purification of a proteolytically activated form Free

Abstract

Summary

A protein of about 800 kDa with trehalose-6-phosphate synthase (TPS) and trehalose-6-phosphate phosphatase (TPP) activity was purified from bakers’ yeast. This TPS/P complex contained 57, 86 and 93 kDa polypeptides. The 86 and 93 kDa polypeptides both appeared to be derived from a polypeptide of at least 115 kDa in the native enzyme. A TPS-activator (a dimer of 58 kDa subunits) was also purified. It decreased the Michaelis constants for both UDP-glucose (three-fold) and glucose 6-phosphate (G6P) (4·5-fold), and increased TPS activity at 5 m-UDP-glucose/10 m-G6P about three-fold. It did not affect TPP activity. The purification of TPS/P included an endogenous proteolytic step that increased TPS activity about three-fold and abolished its requirement for TPS-activator, but did not change TPP activity. This activation was accompanied by a decrease of some 20 kDa in the molecular mass of a cluster of SDS-PAGE bands at about 115 kDa recognized by antiserum to pure TPS/P, but by no change in the 57 kDa band. Phosphate inhibited TPS activity ( about 5 m), but increased TPP activity about six-fold ( about 4 m). Phosphate (6 m) stimulated the synthesis of trehalose from G6P and UDP-glucose and decreased the accumulation of trehalose 6-phosphate.

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© Society for General Microbiology, 1991
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1991-02-01
2024-04-26
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Trehalose-6-phosphate synthase/phosphatase complex from bakers’ yeast: purification of a proteolytically activated form
Microbiology 137, 323 (1991); https://doi.org/10.1099/00221287-137-2-323
/content/journal/micro/10.1099/00221287-137-2-323
/content/journal/micro/10.1099/00221287-137-2-323
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