Purification of Bisphosphoglyceromutase, 2, 3-Bisphosphoglycerate Phosphatase and Phosphoglyceromutase from Human Erythrocytes. Three Enzyme Activities in One Protein

Sasaki, Ryuzo; Ikura, Koji; Sugimoto, Etsuro; Chiba, Hideo

doi:10.1111/j.1432-1033.1975.tb09899.x

Cited by 105 publications

(44 citation statements)

References 34 publications

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“…One of these enzymes is the 2,3-bisphosphoglycerate synthasephosphatase or 2,3-bisphosphoglycerate mutase (EC 5.4.2.4), which is a homodimer of a subunit that possesses great homology with PGM subunits (Sasaki et al, 1975;Kappel and Hass, 1976;Sasaki et al, 1976;Narita et al, 1979). The other two enzyme proteins are heterodimers resulting from the combination of a BPGM subunit with a PGM subunit either type M or type B Pons et al, 1985a).…”

Section: Discussionmentioning

confidence: 99%

Phosphoglycerate mutase, 2,3-bisphosphoglycerate phosphatase and enolase activity and isoenzymes in lung, colon and liver carcinomas

Durany

Joseph²,

Campo³

et al. 1997

Br J Cancer

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SummaryWe have compared the levels of phosphoglycerate mutase, 2,3-bisphosphoglycerate phosphatase and enolase activities and the distribution of their isoenzymes in normal colon, liver and lung tissues, and in colon, liver and lung adenocarcinoma, lung squamous cell carcinoma and lung carcinoid. All tumours presented higher phosphoglycerate mutase and enolase activities and lower 2,3-bisphosphoglycerate phosphatase activity than the normal tissues. No changes were observed in the phosphoglycerate mutase isoenzyme patterns analysed by cellulose acetate electrophoresis. All specimens contained mainly type BB isoenzyme, traces of type MB isoenzyme and no type MM isoenzyme. However, the tumours had decreased levels of 2,3-bisphosphoglycerate mutase and 2,3-bisphosphoglycerate mutase-phosphoglycerate mutase hybrid enzyme. Determined by agarose gel electrophoresis, aa-enolase was the isoenzyme predominant in normal lung, colon and liver tissue, although ay-and yy-enolase were also present in all tissues. In colon, liver and non-endocrine lung tumours, the proportions of ay-and yy-enolase decreased. In contrast, in carcinoid tumours of the lung, the proportions of these isoenzymes increased.Keywords: 2,3-bisphosphoglycerate mutase; 2,3-bisphosphoglycerate phosphatase; enolase; phosphoglycerate mutase; isoenzyme; lung, colon and liver adenocarcinoma; lung squamous cell carcinoma; lung carcinoid Phosphoglycerate mutase (D-phosphoglycerate 2,3-phosphomutase, EC 5.4.2.1, PGM) and enolase (2-phospho-D-glycerate hydrolyase, EC 4.2.1.11) are glycolytic enzymes that catalyse consecutive reversible reactions connecting the two ATP-generating reactions in the glycolytic pathway. PGM catalyses the conversion of 3-phosphoglycerate, product of the first ATP-generating reaction, into 2-phosphoglycerate in the presence of the cofactor 2,3-bisphosphoglycerate. Enolase catalyses the conversion of 2-phosphoglycerate into phosphoenolpyruvate, substrate of the second ATP-generating reaction. In addition to the main mutase activity, PGM possesses collateral 2,3-bisphosphoglycerate synthase or 2,3-bisphosphoglycerate mutase activity (BPGM: 1,3-bisphosphoglycerate + 3-phosphoglycerate -> 3-phosphoglycerate + 2,3-bisphosphoglycerate) and 2,3-bisphosphoglycerate phosphatase activity (BPGP: 2,3-bisphosphoglycerate --3-phosphoglycerate + Pi), which is stimulated by 2-phosphoglycolate (for reviews, see Fothergill-Gilmore and Watson, 1989;Wold, 1971).In mammalian tissues, there are three isoenzymes of PGM, which result from the homodimeric and the heterodimeric combinations of two different subunits coded by separate genes and designated M (muscle) and B (brain). In early fetal life, type BB-PGM is the only form present. During myogenesis, the isoenzyme phenotype undergoes transition, type BB-PGM being replaced by the MM-form, through the MB isoenzyme. In skeletal muscle, there is an almost complete transition from the BB-to the MM-PGM, but in heart muscle complete transition does not occur (Omenn and Cheung, Omenn and Hermodson, 1975;Adamson, 1...

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Section: Discussionmentioning

confidence: 99%

Phosphoglycerate mutase, 2,3-bisphosphoglycerate phosphatase and enolase activity and isoenzymes in lung, colon and liver carcinomas

Durany

Joseph²,

Campo³

et al. 1997

Br J Cancer

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“…The hydrolysis of 2,3-DPG is physiologically stimulated by 2-phosphoglycolate, a normal constituent of red blood cells (4). BPGM also displays a mutase reaction similar to that of the glycolytic enzyme monophosphoglycerate mutase (MPGM, EC 5.4.2.1) which reversibly converts glycerate 3-phosphate (3-PG) to glycerate 2-phosphate (5,6).…”

mentioning

confidence: 99%

Critical Role of Human Bisphosphoglycerate Mutase Cys22 in the Phosphatase Activator-binding Site

Ravel

Craescu

Arous

et al. 1997

Journal of Biological Chemistry

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The enzymatic activities catalyzed by bisphosphoglycerate mutase (BPGM, EC 5.4.2.4) have been shown to occur at a unique active site, with distinct binding sites for diphosphoglycerates and monophosphoglycerates. The physiological phosphatase activator (2-phosphoglycolate) binds to BPGM at an undetermined site. BPGM variants were constructed by site-directed mutagenesis of three amino acid residues in the active site to identify residues specifically involved in the binding of the monophosphoglycerates and 2-phosphoglycolate. Substitution of Cys 22 by functionally conservative residues, Thr or Ser, caused a great decrease in 2-phosphoglycolate-stimulated phosphatase activity and in the K a value of the activator, whereas it caused no change in other catalytic activities or in the K m values of 2,3-diphosphoglycerate (2,3-DPG) and glycerate 3-phosphate (3-PG, EC 1.1.1.12), indicating that Cys 22 is specifically involved either directly or indirectly in 2-phosphoglycolate binding.Kinetic experiments showed that the K a of the cofactor and the K m of 3-PG were affected by the substitution of Ser 23 indicating that this residue is necessary for the fixation of both 3-PG and 2-phosphoglycolate. The R89K variant has previously been shown to have a modified K m value for monophosphoglycerates, however, its affinity for 2-phosphoglycolate is unaltered, suggesting that Arg 89 is specifically involved in monophosphoglycerates binding.CD spectroscopic studies of substrates and cofactor binding showed that 2,3-DPG induced structural modifications of normal and mutated enzymes which could be due to protein phosphorylation. Addition of 2-phosphoglycolate to phosphorylated proteins with normal affinity for the cofactor produced spectra with the same characteristics as unphosphorylated species.In summary, monophosphoglycerates and 2-phosphoglycolate have partially distinct binding sites in human BPGM. The specific implication of the Cys 22 residue in 2-phosphoglycolate binding is of great significance in the design of analogs of therapeutic benefit.

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“…These studies confirm the suggestion of Rosa et al [3] that an enzyme possessing 3-Pglycerate mutase activity also had 2,3-P,glycerate mutase and 2,3-P,glycerate phosphatase activities. The studies of Sasaki et al [4] have been confirmed independently by Hass and Miller [23], Kappel and Hass [5], and Rose and Dube [6].…”

Section: Discussionmentioning

confidence: 75%

“…The ratio between the activities of 2,3-P,glycerate phosphatase and 2,3-P,glycerate mutase were the same in both fractions. These data are interpreted as demonstrating that the 2,3-P2-glycerate phosphatase-mutase of the day-old chick erythrocyte possesses inherent 3-Pglycerate mutase activity as has been shown both for the enzyme from human erythrocytes purified to homogeneity [4,5] and that from equine erythrocytes [6].…”

Section: 3-p2glycerate Phosphatase Activity and 23-p2glycerate Conmentioning

confidence: 76%

“…In this paper we have shown that the red blood cells of the adult chicken contain very low levels of 2,3-P2-glycerate mutase and 2,3-P,glycerate phosphatase activities, but that the activities of these enzymes in the late embryo and hatchling are much higher. Sasaki et al [4] carried out extensive purification of three proteins from human erythrocytes with 3-Pglycerate mutase activity and demonstrated that one possessed quite different kinetic and stability properties when contrasted with the other two which were quite similar. The one enzyme, when purified to apparent homogeneity, displayed three separate activi-ties : 3-Pglycerate mutase, 2,3-P2glycerate mutase and 2,3-P,glycerate phosphatase.…”

Section: Discussionmentioning

confidence: 99%

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Purification and Properties of 2,3-Bisphosphoglycerate Phosphatase-Mutase from Erythrocytes of Day-Old Chicks

1977

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1. Large quantities of 2,3-bisphosphoglycerate accumulate in the red blood cells of the chick embryo during the week prior to hatching; the 2,3-bisphosphoglycerate then abruptly decreases to very small amounts within a few days after hatching.2. The enzyme 2,3-bisphosphoglycerate phosphatase was purified from the red blood cells of the day old chick. The elution profiles for bisphosphoglycerate phosphatase and 2,3-bisphosphoglycerate mutase were identical upon gradient elution from columns of hydroxylapatite and diethylaminoethylcellulose and by gel filtration on Sephadex G-100. At each stage of purification the ratio of phosphatase to mutase activity was the same. It was concluded that, just as in the human erythrocyte, both bisphosphoglycerate phosphatase and 2,3-bisphosphoglycerate mutase activities in chick erythrocytes reside on one protein.3. Bisphosphoglycerate phosphatase has a pH of optimal activity of 7.0. Its activity is stimulated by 2-phosphoglycolate, phosphoenolpyruvate, bisulfite and dithionite and is inhibited by 2-phosphoglycerate, 3-phosphoglycerate, inorganic pyrophosphate and phytic acid. These properties are very similar to those described for bisphosphoglycerate phosphatase purified from human erythrocytes.4. The purified bisphosphoglycerate phosphatase-mutase contained 3-phosphoglycerate mutase activity. Although the 2,3-bisphosphoglycerate mutase and 3-phosphoglycerate mutase activities were nearly equal in the purified enzyme, this protein accounts for a maximum of only 6% of the 3-phosphoglycerate mutase activity in these cells.

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Purification of Bisphosphoglyceromutase, 2, 3-Bisphosphoglycerate Phosphatase and Phosphoglyceromutase from Human Erythrocytes. Three Enzyme Activities in One Protein

Cited by 105 publications

References 34 publications

Phosphoglycerate mutase, 2,3-bisphosphoglycerate phosphatase and enolase activity and isoenzymes in lung, colon and liver carcinomas

Phosphoglycerate mutase, 2,3-bisphosphoglycerate phosphatase and enolase activity and isoenzymes in lung, colon and liver carcinomas

Critical Role of Human Bisphosphoglycerate Mutase Cys22 in the Phosphatase Activator-binding Site

Purification and Properties of 2,3-Bisphosphoglycerate Phosphatase-Mutase from Erythrocytes of Day-Old Chicks

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