Kinetics of triose phosphate isomerase

Burton, Pamela M.; Waley, S. G.

doi:10.1016/0005-2744(68)90028-4

Cited by 47 publications

(14 citation statements)

References 11 publications

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“…No significant differences in the K , values for each of t'he enzymes were observed. The values for the pituitary enzyme are in agreement with those reported by other workers [12,13]. Pituitary triose-phosphate isomerase was active over the measured pH range 6.5 to 8.7 with a pH optimum a t 8.4.…”

Section: Resultssupporting

confidence: 81%

Identification and Characterization of Pituitary Triose‐Phosphate Isomerase

Greenberg

Yen

Bobbitt

1972

European Journal of Biochemistry

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Several pituitary preparations from various species, including two purzed follicle-stimulating hormone preparations, inhibited the ~-glyceraldehyde-3-phosphate dehydrogenase reaction. The active factor in those preparations that caused the apparent inhibition has been identified as an enzyme, triose phosphate isomerase. Using ~-glyceraldehyde-3-phosphate as the substrate, the optimum pH for the isomerization reaction is 8.4. The partially purified enzyme has a specific activity of 34 pmol dihydroxyacetone phosphate isomerized x min-* x mg protein+. The Km of the enzyme for D-glyceraldehyde 3-phosphate is 0.79 mM, and the K m for dihydroxyacetone phosphate is 1.53 mM. Half of the enzyme activity is lost in 7 min at 55 "C. The molecular weight of the enzyme is estimated to be 56000. The electrophoretic mobility of the enzyme differs from tha.t of the muscle enzyme supplied by Sigma indicating a difference in amino acid composition between the two enzymes. Electrophoresis revealed the presence of two to three isozymes of the pituitary triose phosphate isomerase.Two preparations of follicle-stimulating hormone, supplied by The National Institute of Arthritis and Metabolic Disease, inhibited the reaction of D-glyceraldehyde-3-phosphate dehydrogenase using glyceraldehyde-3-phosphate as the substrate [l]. These two preparations were of ovine (lot #S-4) and porcine (lot # P-1) origin. Another ovine preparation (lot #S-7) did not inhibit this reaction [l]. We thus concluded that the inhibitor was not follicle-stimulating hormone but a contaminant [l]. We also concluded that the inhibitor was not one of the known pituitary hormones we had tested, since none of them inhibited the ~-glyceraldehyde-3-phosphate dehydrogenase reaction El].Bornstein and his associates have reported the isolation of a factor, InG, from a pH-2 hydrolyzate of ovine growth hormone that inhibited D-glyceraldehyde-3-phosphate dehydrogenase and possessed other biological properties [2,3]. The amino acid sequence at both the amino terminal and carboxyl terminal of InG suggested that InG was a peptide from growth hormone that consisted of residues 164 to 188 [4]. A peptide synthesized according to the proposed amino acid sequence of InG had all the biological activities of the isolated InG [a].

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Section: Resultssupporting

confidence: 81%

Identification and Characterization of Pituitary Triose‐Phosphate Isomerase

Greenberg

Yen

Bobbitt

1972

European Journal of Biochemistry

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“…absorption of the enzyme was altered, and there were crystallographic changes that suggested changes in conformation of the enzyme. Glycerol 3-phosphate, a substrate analogue that is a competitive inhibitor (Burton & Waley, 1968a), had similar effects on the u.v. absorption (Johnson & Wolfenden, 1970).…”

Section: (Received 1 September 1978)mentioning

confidence: 93%

Spectrophotometric studies on the interaction between triose phosphate isomerase and inhibitors

Jones¹,

Waley²

1979

Biochemical Journal

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The binding of ligands to chicken muscle triose phosphate isomerase was studied. Changes in u.v. absorbance of the enzyme were used to measure binding, and the dissociation constant was determined over a range of pH values. The ligands were 2-phosphoglycollate and rac-glycerol 3-phosphate (only the D-isomer, sn-glycerol 1-phosphate, binds appreciably). Non-linear regression was used to fit calculated curves to the experimental points and hence to compare different models. Both active sites in the dimeric enzyme probably bound 2-phosphoglycollate, without any interaction between the sites. The results of crystallographic analysis [Phillips, Rivers, Sternberg, Thornton & Wilson (1977) Biochem. Soc. Trans. 5,[642][643][644][645][646][647], and experiments on the 1H, 13C and 31P n.m.r. of enzyme or 2-phosphoglycollate were combined with the present results to provide the basis for a model in which binding depends on glutamic acid-165 being protonated and on the ligand being fully ionized; additionally, binding affects the ionization of one histidine residue (probably histidine-100). The binding of the glycerol 3-phosphate, on the other hand, was independent of pH over the range pH 6.5-8.5 but decreased at lower pH values. This is explained on a model in which the binding of the monoanion of the ligand is markedly affected by the protonation of a residue in the enzyme, but the binding of the dianion is only slightly affected by this ionization.

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“…4 must be corrected for the effect of arsenate on the isomerase reaction (an effect first noted by Burton & Waley, 1968b). The K1 for competitive inhibition by arsenate of the reaction with dihydroxyacetone phosphate as substrate, under the assay conditions used here (I0.1, 30°C, pH7.5) was found to be 11.1 mm [Burton & Waley (1968b) report 5.5mM at 25°C for the rabbit muscle enzyme]. The apparent Km for dihydroxyacetone phosphate (1.5mM) was corrected by using this value, to give 0.97mM.…”

Section: Kinetics Of the Triose Phosphate Isomerase Reactionmentioning

confidence: 99%

Specificity and kinetics of triose phosphate isomerase from chicken muscle

Putman

Coulson

Farley

et al. 1972

Biochemical Journal

109

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The isolation of crystalline triose phosphate isomerase from chicken breast muscle is described. The values of k(cat.) and K(m) for the reaction in each direction were determined from experiments over wide substrate-concentration ranges, and the reactions were shown to obey simple Michaelis-Menten kinetics. With d-glyceraldehyde 3-phosphate as substrate, k(cat.) is 2.56x10(5)min(-1) and K(m) is 0.47mm; with dihydroxyacetone phosphate as substrate, k(cat.) is 2.59x10(4)min(-1) and K(m) is 0.97mm. The enzyme-catalysed exchange of the methyl hydrogen atoms of the ;virtual substrate' monohydroxyacetone phosphate with solvent (2)H(2)O or (3)H(2)O was shown. This exchange is about 10(4)-fold slower than the corresponding exchange of the C-3 hydrogen of dihydroxyacetone phosphate. The other deoxy substrate, 3-hydroxypropionaldehyde phosphate, was synthesized, but is too unstable in aqueous solution for analogous proton-exchange reactions to be studied.

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Kinetics of triose phosphate isomerase

Cited by 47 publications

References 11 publications

Identification and Characterization of Pituitary Triose‐Phosphate Isomerase

Identification and Characterization of Pituitary Triose‐Phosphate Isomerase

Spectrophotometric studies on the interaction between triose phosphate isomerase and inhibitors

Specificity and kinetics of triose phosphate isomerase from chicken muscle

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