Nature of the subunits of the 6-phosphofructo-1-kinase isoenzymes from rat tissues

Dunaway, George; Kasten, Tom

doi:10.1042/bj2420667

Cited by 65 publications

(52 citation statements)

References 24 publications

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“…This molecular mass value is in the upper limit of those reported for other eukaryotic PFruKs. Thus it is lower than those determined for the two different subunits of yeast PFruK (108 and 105 kDa for the a and subunits, respectively [5]), similar to that of the Ascaris suum enzyme [30], and above those of the three subunit types of mammalian PFruKs (80, 85 and 87.5 kDa for the L, M and C subunits, respectively [31]) and the enzymes from trout muscle (76.5 kDa) [32], Fusciola hepatica (83 kDa) [33] and Trypanosoma brucei (49 kDa) [34]. When the slime mold PFruK was subjected to gel filtration on Superose 6B, the enzyme eluted as a single peak (Fig.…”

Section: Results Purificationmentioning

confidence: 67%

“…The procedure has been performed about 15 times with similar results and provided us with fairly good recovery, in spite of the small activity of the cell extract and the number of steps involved (Table 1). In contrast to PFruKs from yeast [ S ] and mammalian cells [31], which contain two and three types of subunits, respectively, the enzyme from the slime mold is composed of a single type of polypeptide chain of 95 kDa, the native enzyme being a homotetramer of 382 kDa. Gel filtration studies suggest (Fig.…”

Section: Discussionmentioning

confidence: 99%

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Purification and Properties of Phosphofructokinase from Dictyostelium Discoideum

Martı́nez-Costa

Estévez

Sánchez

et al. 1994

European Journal of Biochemistry

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Phosphofructokinase (PFruK) from the slime mold Dictyostelium discoideum has been purified to homogeneity over 15000-fold with a 29% yield. Sodium dodecyl sulfate/polyacrylamide gel electrophoresis of the final preparation revealed a single band of 95 kDa. The native molecular mass was determined by gel filtration to be 382 kDa, indicating that the enzyme is a homotetramer. An antibody raised in rabbits against the 95-kDa band immunoprecipitated PFruK activity while it did not react with the enzyme from yeast and mammalian cells. The apparent PI was 6.8 and the pH optimum was 7.6. The enzyme had an activation energy (EJ of 29

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Section: Results Purificationmentioning

confidence: 67%

Section: Discussionmentioning

confidence: 99%

Purification and Properties of Phosphofructokinase from Dictyostelium Discoideum

Martı́nez-Costa

Estévez

Sánchez

et al. 1994

European Journal of Biochemistry

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“…There are three main isoforms of PFK: PFK-M (alternatively known as PFK-A), PFK-L (alternatively known as PFK-B), and PFK-C. PFK-M the only isoform of PFK found in skeletal muscle and diaphragm (31). PFK-L is the major isoform of PFK expressed in liver (31).…”

Section: Discussionmentioning

confidence: 99%

“…1, proteins of 45, 53, 60, and 65 kDa leached from the nNOS-Affi-Gel buffer control column (lanes 1 and 4). Additionally, a number of nonspecific proteins (31,33,35,39, and 50 kDa) eluted from both the nNOS-and GST-Affi-Gel columns (lanes 2, 3, 5, and 6). These proteins either bind to GST or nonspecifically bind to Affi-Gel beads.…”

Section: Isolation Of An 80-kda Protein Thatmentioning

confidence: 99%

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Interaction of Neuronal Nitric-oxide Synthase and Phosphofructokinase-M

Firestein

Bredt

1999

Journal of Biological Chemistry

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Neurons that express neuronal nitric-oxide synthase (nNOS) are resistant to NO-induced neurotoxicity; however, the mechanism by which these neurons are protected is not clear. To identify proteins possibly involved in this process, we performed affinity chromatography with the nNOS PDZ domain, a N-terminal motif that mediates protein interactions. Using this method to fractionate soluble tissue extracts, we identified the muscle isoform of phosphofructokinase (PFK-M) as a protein that binds to nNOS both in brain and skeletal muscle. PFK-M interacts with the PDZ domain of nNOS, and nNOS-PFK-M binding can be competed by peptides that bind to the PDZ domain of nNOS. We found that nNOS is significantly associated with PFK-M in skeletal muscle because nNOS can be immunodepleted from cytosolic skeletal muscle extracts using an antibody directed against PFK-M. In brain, nNOS and PFK-M are both enriched in synaptosomes, and specifically, in the synaptic vesicle fraction, where they can interact. At the cellular level, PFK-M is enriched in neurons that express nNOS protein. As fructose-1,6-bisphosphate, the product of PFK activity, is neuroprotective, the interaction of nNOS and PFK may contribute to neuroprotection of nNOS positive cells.Nitric oxide, derived from neuronal nitric-oxide synthase (nNOS), 1 has important physiological functions in the nervous system (1, 2). In the peripheral nervous system, NO mediates certain actions of autonomic motor neurons on smooth muscle and thereby NO regulates intestinal peristalsis and penile erection. In the central nervous system, NO does not primarily act on smooth muscle but instead modulates synaptic transmission associated with N-methyl-D-aspartate (NMDA) type glutamate receptors. By regulating synaptic plasticity, NO can influence hippocampal long term potentiation (3) as well as aspects of learning (4) and memory (5).Although small amounts of NO mediate physiological signaling, excess NO production can cause tissue injury (6). The primary stimulus for NO synthesis in central neurons is activation of NMDA receptors (2). Overactivity of NMDA receptors is implicated in numerous neurodegenerative processes including stroke, Huntington's chorea, and amyotrophic lateral sclerosis (7). A role for NO in NMDA-mediated degeneration was first suggested by experiments showing neuroprotection by NO synthase antagonists (8). Definitive evidence that nNOS mediates brain injury derives from studies of nNOS knockout mice, which are strikingly resistant to excitotoxicity following focal cerebral ischemia (9).NO can mediate neurotoxicity by inhibiting metabolic pathways and causing cellular energy depletion, which is a hallmark of ischemic injury-induced neuronal death. NO erodes energy stores by reacting with certain metabolic enzymes that contain heme-iron prosthetic groups, iron-sulfur clusters, or reactive thiols (10). Through these reactions, NO can inhibit glycolysis by reacting with cis-aconitase and can attenuate oxidative phosphorylation by inhibiting mitochondrial iron-sulfur enzy...

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Phosphofructokinase in the rat nervous system: Regional differences in activity and characteristics of axonal transport

Oblinger

Foe

Kwiatkowska

et al. 1988

J of Neuroscience Research

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The distribution of phosphofructokinase (PFK) in gray and white matter regions of the rat nervous system was evaluated. Determinations of PFK activity revealed that cell body enriched regions (sensorimotor cortex) had a significantly higher level of activity than axonal regions (sciatic nerve, dorsal roots, and optic nerve). The level of PFK activity was also significantly higher in central axons (optic nerve) than in peripheral axons (sciatic nerve). Differences in PFK activity could be largely attributed to differences in tissue content of the enzyme rather than to differences in the types of PFK isozymes present. Cortex contained significantly larger amounts of PFK relative to total protein than did peripheral nerve. However, purification of PFK revealed that all three of the PFK isozymes, C (86 kd), A (84 kd), and B (80 kd), were present in both cortex and sciatic nerve. Both SDS/PAGE and immunoblotting studies using PFK isozyme-specific antibodies demonstrated that the relative proportions of the three PFK isozymes were similar in cell body and axonal regions of the nervous system. The PFK-C and PFK-A isozymes each comprised about half the total and only small amounts of the PFK-B isozyme were present in both regions. However, immunoprecipitation experiments suggested that quantitatively different proportions of the possible PFK hybrids (tetramers) may be distributed between axonal and cell body regions. The transport of PFK was examined in this study and PFK was identified in slow component b (SCb) of axonal transport. SCb moves at a rate of 2-4 mm/day in rat axons and is known to contain several other enzymes of intermediary metabolism as well as actin. The finding that PFK, the rate limiting enzyme in glycolysis, is present in SCb lends support to the hypothesis that glycolytic enzymes are not freely diffusing proteins in axons but, instead, are present as organized assemblies that have long-term, yet flexible, associations with structural elements of the cytoplasm.

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Nature of the subunits of the 6-phosphofructo-1-kinase isoenzymes from rat tissues

Cited by 65 publications

References 24 publications

Purification and Properties of Phosphofructokinase from Dictyostelium Discoideum

Purification and Properties of Phosphofructokinase from Dictyostelium Discoideum

Interaction of Neuronal Nitric-oxide Synthase and Phosphofructokinase-M

Phosphofructokinase in the rat nervous system: Regional differences in activity and characteristics of axonal transport

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