Interaction of muscle glycolytic enzymes with thin filament proteins

Bronstein, W. W.; Knull, Harvey R.

doi:10.1139/o81-069

Cited by 109 publications

(60 citation statements)

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“…Numerous studies have demonstrated interactions between the cytoskeleton and glycolytic enzymes, especially GAPDH, aldolase and PK. [25][26][27][28][29][30][31][32][33][34][35][36][37][38] In this study, quantities of these glycolytic enzymes, as well as actin, tubulin and tropomyosin, were sufficient in U87 pseudopodia to be detected with Figure 4 DeCyder ratios Coomassie blue staining, thus establishing the potential for the energy-generating portion of the glycolytic pathway to energize cytoskeletal proteins for migration within U87 pseudopodia. Polymerization of actin and tubulin shares a similar use of energy from nucleotide triphosphate hydrolysis.…”

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confidence: 63%

Proteomic characterization of harvested pseudopodia with differential gel electrophoresis and specific antibodies

Beckner

Chen

et al. 2005

Laboratory Investigation

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Malignant gliomas (astrocytomas) are lethal tumors that invade the brain. Invasive cell migration is initiated by extension of pseudopodia into interstitial spaces. In this study, U87 glioma cells formed pseudopodia in vitro as cells pushed through 3 lm pores of polycarbonate membranes. Harvesting pseudopodia in a novel two-step method provided material for proteomic analysis. Differences in the protein profiles of pseudopodia and whole cells were found using differential gel electrophoresis (DIGE) and immunoblotting. Proteins from twodimensional (2D) gels with M R 's of 20-100 kDa and pI's of 3.0-10.0 were identified by peptide mass fingerprinting analysis using mass spectrometry. For DIGE, lysates of pseudopodia and whole cells were each labeled with electrophilic forms of fluorescent dyes, Cy3 or Cy5, and analyzed as mixtures. Analysis was repeated with reciprocal labeling. Differences in protein distributions were detected by manual inspection and computer analysis. Topographical digital maps of the scanned gels were used for algorithmic spot matching, normalization of background, quantifying spot differences, and elimination of artifacts. Pseudopodial proteins in Coomassie-stained 2D gels included isoforms of glycolytic enzymes as the largest group, seven of 24 proteins. Peptide mass fingerprint analysis of DIGE gels demonstrated increased isoforms of annexin (Anx) I, AnxII, enolase, pyruvate kinase, and aldolase, and decreased mitochondrial manganese superoxide dismutase and transketolase in pseudopodia. Specific antibodies showed restricted immunoreactivity of the hepatocyte growth factor (HGF) a chain to pseudopodia, indicating localization of its active form. Met (the HGF receptor), actin, and total AnxI were increased in pseudopodial lysates on immunoblots. Increased constituents of the pseudopodial proteome in glioma cells, identified in this study as actin, HGF, Met, and isoforms of AnxI, AnxII, and several glycolytic enzymes, represent therapeutic targets to consider for suppression of tumor invasion.

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confidence: 63%

Proteomic characterization of harvested pseudopodia with differential gel electrophoresis and specific antibodies

Beckner

Chen

et al. 2005

Laboratory Investigation

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“…Aldolase interacts with calmodulin (20,75) and phospholipase D 2 (21) and therefore may modulate signal transduction pathways. Aldolase is also an actin-binding protein that exhibits a dynamic interaction with the cytoskeleton (22,23). Importantly, aldolase interacts with the H ϩ -ATPase (25,26) and GLUT4 (27), two proteins regulated by intracellular trafficking.…”

Section: Discussionmentioning

confidence: 99%

“…Aldolase Binding Reduces NKCC2 Surface Expression-Aldolase is a protein associated with the actin cytoskeleton and can cross-link actin fibers (22,23). Because actin cytoskeleton has been implicated in the regulation of Na-K-Cl co-transport in mouse kidney cultured TAL cells (58), we therefore sought to study the effect of aldolase binding on NKCC2 surface expression.…”

Section: Endogenous Aldolase Protein Interacts and Co-localizes With mentioning

confidence: 99%

NKCC2 Surface Expression in Mammalian Cells

Benziane

Demaretz

Defontaine

et al. 2007

Journal of Biological Chemistry

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Apical bumetanide-sensitive Na؉ -K ؉ -2Cl ؊ co-transporter, termed NKCC2, is the major salt transport pathway in kidney thick ascending limb. NKCC2 surface expression is subject to regulation by intracellular protein trafficking. However, the protein partners involved in the intracellular trafficking of NKCC2 remain unknown. Moreover, studies aimed at understanding the post-translational regulation of NKCC2 have been hampered by the difficulty to express NKCC2 protein in mammalian cells. Here we were able to express NKCC2 protein in renal epithelial cells by tagging its N-terminal domain. To gain insights into the regulation of NKCC2 trafficking, we screened for interaction partners of NKCC2 with the yeast two-hybrid system, using the C-terminal tail of NKCC2 as bait. Aldolase B was identified as a dominant and novel interacting protein. Real time PCR on renal microdissected tubules demonstrated the expression of aldolase B in the thick ascending limb. Co-immunoprecipitation and co-immunolocalization experiments confirmed NKCC2-aldolase interaction in renal cells. Biotinylation assays showed that aldolase co-expression reduces NKCC2 surface expression. In the presence of aldolase substrate, fructose 1,6-bisphosphate, aldolase binding was disrupted, and aldolase co-expression had no further effect on the cell surface level of NKCC2. Finally, functional studies demonstrated that aldolase-induced down-regulation of NKCC2 at the plasma membrane was associated with a decrease in its transport activity. In summary, we identified aldolase B as a novel NKCC2 binding partner that plays a key role in the modulation of NKCC2 surface expression, thereby revealing a new regulatory mechanism governing the co-transporter intracellular trafficking. Furthermore, NKCC2 protein expression in mammalian cells and its regulation by protein-protein interactions, described here, may open new and important avenues in studying the cell biology and post-transcriptional regulation of the co-transporter.

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“…The choice of a 0.05M ionic strength is in accordance with a variety of experimental investigations. [2][3][4] This choice also enhanced the interaction making it easier to observe statistically. The 0.05M ionic strength increased the number of bimolecular complexes and allowed for a better visualization of the electrostatic field.…”

Section: Charge Assignments and Epsmentioning

confidence: 99%

“…For example, the glycolytic enzymes fructose-1,6-bisphosphate aldolase (aldolase) and glyceraldehyde-3-phosphate dehydrogenase (GADPH)) have been shown to interact with cytoskeletal proteins, especially F-actin in human, 1 rabbit, 2 fish, 3 and possibly Drosophila. 4 These interactions have been studied by a variety of experimental methods such as affinity chromatography, 2 histochemical observations, 3 copelleting, 5 and co-electrophoresis. 6 These interactions have been postulated to be electrostatic in nature due to their dependence on pH, ionic strength, monovalent (K + , Na + ) or divalent (Ca 2+ , Mg 2+ ) cations, as well as metabolites (ATP, ADP).…”

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confidence: 99%

Theoretical study of interactions between muscle aldolase and F‐actin: Insight into different species

Forlemu¹,

Waingeh²,

Ouporov³

et al. 2006

Biopolymers

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Interactions of the glycolytic enzyme, fructose‐1,6‐bisphosphate aldolase (aldolase), with F‐actin may be one mechanism for the colocalization of glycolytic enzymes. Examination of these interactions in different animal species tests this hypothesis by observing whether binding sites are conserved across species. Brownian dynamics (BD) simulations provide descriptions of such protein–protein interactions with the muscle isoforms of zebra fish and human aldolase. The results are compared with previous results obtained for rabbit muscle and yeast. The aldolase binding groove previously determined in rabbit muscle is conserved in both the human and fish muscle isoforms. The nonspecific radial free energies of interaction are similar with fish being slightly weaker than human and rabbit: human, −2.27 ± 0.05 kcal/mol; rabbit, −2.0 ± 0.04 kcal/mol; and fish, −1.5 ± 0.03 kcal/mol. BD results show a large Boltzmann population of complexes formed around the A/D and B/C grooves of aldolase with the most feasible binding mode comprising two aldolase subunits to subdomain I region of the actin subunits. These results show that the location of the important residues and binding site for fish and human aldolase is very similar to that in rabbit and that in different animals the binding site is conserved. This suggests that the binding interaction between aldolase and F‐actin is general in animal muscles and is rendered possible and energetically favorable through the conservation of this binding site. © 2006 Wiley Periodicals, Inc. Biopolymers 85: 60–71, 2007.This article was originally published online as an accepted preprint. The “Published Online” date corresponds to the preprint version. You can request a copy of the preprint by emailing the Biopolymers editorial office at biopolymers@wiley.com

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Interaction of muscle glycolytic enzymes with thin filament proteins

Cited by 109 publications

References 0 publications

Proteomic characterization of harvested pseudopodia with differential gel electrophoresis and specific antibodies

Proteomic characterization of harvested pseudopodia with differential gel electrophoresis and specific antibodies

NKCC2 Surface Expression in Mammalian Cells

Theoretical study of interactions between muscle aldolase and F‐actin: Insight into different species

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