Ligand-induced conformational states of rabbit liver fructose 1,6-bisphosphatase as revealed by digestion with subtilisin

Pontremoli, S.; Melloni, Edon; Balestrero, F.; Flora, Antonio De; Horecker, B.L.

doi:10.1016/0003-9861(73)90363-9

Cited by 24 publications

(5 citation statements)

References 15 publications

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“…This was in contrast to the result reported in Fig. 3A We have previously shown that conformation changes in rabbit liver Fru-P2ase can be monitored by measuring its susceptibility to limited proteolysis by subtilisin (26). On this basis, aldolase appears to induce a more resistant conformation in Fru-P2ase.…”

Section: Gelcontrasting

confidence: 56%

Evidence for formation of a rabbit liver aldolase--rabbit liver fructose-1,6-bisphosphatase complex.

MacGregor¹,

Singh²,

Davoust³

et al. 1980

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

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The ability of rabbit liver aldolase (D-fructose-1,6-bisphosphate D-glyceraldehyde-3-phosphatate-lyase, EC 4.1.2.13) and rabbit liver fructose-1,6-bisphosphatase (FruP2ase; D-fructose-1,6-bisphosphate 1-phosphohydrolase, EC 3.1.3.11) to partition into the gel phase of Ultrogel AcA 34 is decreased in a mixture of the two enzymes. Titration experiments indicate that a 1:1 complex is formed. The value for the distribution coefficient of the complex corresponds to a molecular mass of 300,000 daltons, the value expected for a dimer containing one mole of each enzyme protein. Complex formation was not observed when either liver enzyme was replaced by the corresponding isozyme from rabbit muscle. The susceptibility of liver Fru-Pease to limited proteolysis by subtilisin was reduced in the presence of liver aldolase, but not when the latter was replaced by muscle aldolase, suggesting that the conformation of Fru-P2ase is altered in the complex. Limited proteolysis of liver aldolase abolishes its ability both to form theterodimer and to protect Fru-P2ase from modification by subtilisin.In mammalian liver fructose-1,6-bisphosphate aldolase (D-fructose-1,6-bisphosphate D-glyceraldehyde-3-phosphate-lyase, EC 4.1.2.13) and fructose-1,6-bisphosphatase fructose-1,6-bisphosphate 1-phosphohydrolase, EC 3.1.3.11) catalyze successive reactions in gluconeogenesis. Both are found in the cytosol and, like other enzymes of glycolysis and gluconeogenesis, are considered to exist and function as independent entities. In recent years, however, evidence has been accumulating for loose interactions between cytosolic proteins and between these proteins and cellular matrices. Arnold et al. (1,2) have reported that several glycolytic enzymes of muscle, including aldolase and glyceraldehyde-3-phosphate dehydrogenase, tend to associate with muscle actin, and Clarke and Masters (3) have examined these interactions in detail under conditions of physiological ionic strength. In mammalian erythrocytes both aldolase (4, 5) and glyceraldehyde-3-phosphate dehydrogenase (6) bind to the major membrane polypeptide (band 3 polypeptide), suggesting that these enzymes may function as a membrane-bound complex. Kinetic and physicochemical evidence for an interaction between aldolase and glyceraldehyde-3-phosphate dehydrogenase purified from rabbit muscle has been reported by Keleti and his coworkers (7,8).It has been proposed (3, 9, 10) that weak interactions between soluble enzymes may play a role in metabolism, either by promoting substrate channeling, in the case of enzymes that catalyze successive reactions, or by the induction of conformation changes that alter their catalytic or regulatory properThe publication costs of this article were defrayed in part by page charge payment. This article must therefore be hereby marked "advertisement" in accordance with 18 U. S. C. §1734 solely to indicate this fact. 3889 ties. Although direct evidence for subtle interactions has been difficult to obtain, the kinetic evidence for substrate channeling repo...

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Section: Gelcontrasting

confidence: 56%

Evidence for formation of a rabbit liver aldolase--rabbit liver fructose-1,6-bisphosphatase complex.

MacGregor¹,

Singh²,

Davoust³

et al. 1980

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

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“…In previous studies the enzyme was found to be susceptible to limited proteolysis during isolation (Geller et al 1971;Traniello et al 1971;Sarngadharan and Pogell 1972) or by treatment of the purified enzyme with subtilisin (Geller et al 1971;Pontremoli et al 1973) to yield an S-peptide of 60 amino acids that could be dissociated in disaggregating solvents. The amino acid sequence of this S-peptide has been established for the enzyme from rabbit liver (EI-Dorry et al 1977), sheep liver (Fisher and Thompson 1980) and pig kidney (Marcus et al 1981) and there was striking homology between the different sequences (Fisher and Thompson 1980;Macgregor et al 1982).…”

Section: Introductionmentioning

confidence: 99%

Amino Acid Sequence Studies on Sheep Liver Fructose-bisphosphatase. II The Complete Sequence

Wk¹,

Eo²

1983

Aust. Jnl. Of Bio. Sci.

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The cyanogen bromide fragments of S-carboxymethylated fructose-bisphosphatase were purified. The amino acid sequences of the small fragments were determined by the dansyl-Edman method. The large fragments were subjected to proteolytic digestion to give smaller peptides more amenable for purification and sequencing by similar methods. Enzyme digests of the S-carboxymethylated enzyme gave overlap peptides containing the methionine residues.In conjunction with the amino acid sequence of the 60-residue N-terminal fragment previously determined on the S-peptide released by limited proteolysis with subtilisin the complete sequence of 336 residues was deduced. The sequence has been compared with the 335 residue sequence of pig kidney fructose-bisphosphatase and some areas of sequence for rabbit liver enzyme. The strong homology previously noted for the S-peptide sequences is maintained for the complete enzyme with only 34 changes in 336 residues when comparing the pig and sheep enzymes.

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“…The native enzymes from rabbit liver and kidney are characterized by having optimum activity at neutral pH, and each is composed of four subunits, whose molecular weight is approximately 36,000 (6,7). Each subunit contains a single tryptophan residue (7,9,10), located in the NHrterminal region of the molecule (11). Exposure of the purified native enzyme to proteolytic enzymes, or to lysosomal fractions, has been shown to cause marked changes in the catalytic properties of the enzyme (9,12,13), notably a shift from a neutral to an alkaline pH optimum.…”

mentioning

confidence: 99%

Changes in Activity and Molecular Properties of Fructose 1,6-Bisphosphatase During Fasting and Refeeding

Pontremoli¹,

Melloni²,

Salamino³

et al. 1974

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

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During prolonged starvation, fructose 1,6-bisphosphatase (EC 3.1.3.11) activity in rabbit liver and kidney shows a transient decrease during the first 36 hr., before rising at 96 hr to levels severalfold higher than those found in the livers of fed animals. Proteolytic activity appears in the 105,000 X g supernatant fraction within several hours of starvation, and continues to increase during the entire 96-hr period. On refeeding, the activities return to nearly the control levels within 24 hr. The catalytic properties of fructose 1,6-bisphosphatase isolated from the livers of fasted rabbits are similar to those of the enzyme from fed animals, but its structure is modified, since it no longer contains the single tryptophan residue located near the NH2-terminus in the native enzyme. Thus this tryptophan residue is not required for the neutral pH optimum. The structural changes and the transient decrease in activity may be related to the observed increase in "free" proteolytic activity.

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Ligand-induced conformational states of rabbit liver fructose 1,6-bisphosphatase as revealed by digestion with subtilisin

Cited by 24 publications

References 15 publications

Evidence for formation of a rabbit liver aldolase--rabbit liver fructose-1,6-bisphosphatase complex.

Evidence for formation of a rabbit liver aldolase--rabbit liver fructose-1,6-bisphosphatase complex.

Amino Acid Sequence Studies on Sheep Liver Fructose-bisphosphatase. II The Complete Sequence

Changes in Activity and Molecular Properties of Fructose 1,6-Bisphosphatase During Fasting and Refeeding

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