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Data Brief. 2017 October; 14: 329–336.
Published online 2017 July 28. doi:  10.1016/j.dib.2017.07.069
PMCID: PMC5547241

Supportive data on the regulation of GLUT4 activity by 3-O-methyl-D-glucose

Abstract

The data presented in this article are related to the research article entitled “Regulation of GLUT4 activity in myotubes by 3-O-methyl-D-glucose” (Shamni et al., 2017) [1]. These data show that the experimental procedures used to analyze the effects of 3-O-methyl-D-glucose (MeGlc) on the rate of hexose transport into myotubes were valid and controlled. The stimulatory effect of MeGlc was limited to glucose transporter 4 (GLUT4) and was independent of ambient glucose and protein synthesis. Cornish-Bowden kinetic analysis of uptake data revealed that MeGlc attenuated indinavir-induced inhibition of hexose transport in a competitive manner.

Keywords: Glucose transporter 4, Indinavir, Intrinsic activity, 3-O-methyl glucose, Myotubes

Specifications Table

Table thumbnail

Value of the data

  • • We have shown that MeGlc augments the intrinsic activity of GLUT4 in myotubes [1]. The data here show that the assays used to analyze the effect of the MeGlc on GLUT4 activity were valid and reproducible and that the experiments were well-controlled.
  • • The activity of GLUT1, in contrary to GLUT4, was not modified in the presence of MeGlc.
  • • The effect of MeGlc on GLUT4 was stereospecific.
  • • MeGlc reduced indinavir-induced inhibition of hexose transport by GLUT4 in a competitive manner.

1. Data

The data presented here are supportive to the data presented in [1] with no duplications or overlap. The data in Fig. 1 show that repetitive exposure of L6 myotubes to MeGlc augmented the rate of hexose transport into L6 myotubes that were maintained at 25 mM glucose. Fig. 2 shows that MeGlc stimulated hexose uptake in L6 myotubes maintained at 5 mM glucose and that these effects were similar to those observed under 25 mM glucose (Fig. 2 in [1]). No such stimulatory effects of MeGlc were evident in GLUT1 expressing vascular cells (Fig. 3). Analyses of [3H]MeGlc transport into wild-type L6 myotubes (Fig. 4) and of [3H] dGlc uptake into L6 myotubes expressing GLUT4myc (Fig. 5, Fig. 6) confirm the validity and suitability of the assays used. Inhibition of protein synthesis with cycloheximide did not interfere with MeGlc effects (Fig. 7). MeGlc exerted its effects also in the presence of glucose in the uptake assay (Fig. 8). Unlike MeGlc, its analog 1-α-methylglucose (1-α-MeGlc) failed to modulate the hexose transport system (Fig. 9). Finally, Cornish-Bowden analysis of the uptake data shows that MeGlc attenuated indinavir-induced inhibition of hexose transport in a competitive manner (Fig. 10).

Fig. 1
Repetitive MeGlc stimulations. L6 myotubes that had been maintained at 25 mM glucose for 24 h were washed and incubated in glucose-free α-Minimum Essential Medium (MEM) supplemented with 20 mM MeGlc (red squares) or 20 mM ...
Fig. 2
(A–C): MeGlc augments the rate of [3H]dGlc uptake in L6 myotubes at 5 mM glucose. A, Time-course analysis: Myotube cultures that had been maintained at 5 mM glucose for 24 h were washed with PBS and received glucose-free ...
Fig. 3
Lack of effect of MeGlc on hexose transport in vascular cells. Primary cultures of bovine aortic endothelial cells (black bars) and smooth muscle cells (white bars) were prepared as describe [2] and maintained with 5 mM glucose in the culture ...
Fig. 4
Time-course analysis of MeGlc transport. L6 myotubes were exposed to 20 mM MeGlc (red squares) or 20 mM L-glucose (blue circles) for 1 h and then were taken for the [3H]MeGlc transport assay at room temperature (left panel) or ...
Fig. 5
MeGlc augments the rate of [3H]dGlc uptake in L6-GLUT4myc myotubes. L6-GLUT4myc Myotube cultures that had been maintained at 25 mM glucose for 24 h were washed with PBS and received glucose-free α-MEM or glucose-free α-MEM ...
Fig. 6
MeGlc stimulatory effect on dGlc uptake is preserved in L6- GLUT4myc myotubes under conditions resembling the conditions of the colorimetric assay. L6-GLUT4myc-expressing myotubes were treated as described in the legend to Fig. 5. The uptake assay was ...
Fig. 7
MeGlc augments hexose uptake in the presence of cycloheximide. L6 myotubes that had been maintained at 25 mM glucose for 24 h were exposed to 1 µM cycloheximide (CHI, white bars) or the vehicle (DMSO, black bars) during ...
Fig. 8
MeGlc stimulates hexose transport in the presence of glucose. L6 myotubes that had been maintained at 25 mM glucose for 24 h were supplemented with the indicated concentrations of MeGlc (squares) or L-Glc (triangles), incubated for an ...
Fig. 9
lack of effect of 1-α-Methyl-D-glucose on hexose transport in L6 myotubes. A, Competition assay: The rate of [3H]dGlc uptake was measured in L6 myotube cultures in the presence of 20 mM of non-radioactive Glc, MeGlc, L-Glc or 1-α-MeGlc ...
Fig. 10:
Kinetic analyses of indinavir-dependent inhibition of hexose uptake in L-Glc- and MeGlc-treated L6 myotubes. A, Dose-response analysis of indinavir-induced inhibition of [3H]dGlc uptake in the presence of increasing concentrations of dGlc in the uptake ...

2. Experimental design, materials and methods

2.1. Cells and treatments

Wild-type L6 myotubes, L6 myotubes expressing GLUT4myc and primary cultures of bovine aortic endothelial and smooth muscle cells were treated as described and then taken for standard [3H]dGlc uptake or [3H]MeGlc transport assays.

Funding information

The authors would like to acknowledge support from the Hebrew University Applied Research Fund A (2011), the Baby Seed Fund of the Yissum Research Development Company of the Hebrew University of Jerusalem (2011), and the Alex Grass Center for Drug Design and Synthesis at the Hebrew University (2013). O.S. and G.C. received fellowships from the Hebrew University Center for Diabetes Research.

Acknowledgements

These data were used in part in the doctoral thesis Ofer Shamni submitted to the Senate of the Hebrew University of Jerusalem.

Footnotes

Transparency documentTransparency data document associated with this article can be found in the online version at http://dx.doi.org/10.1016/j.dib.2017.07.069.

Transparency document. Supplementary material

Transparency document

.

References

1. Shamni O., Cohen G., Gruzman A., Zaid H., Klip A., Cerasi E., Sasson S. Regulation of GLUT4 activity in myotubes by 3-O-methyl-D-glucose. Biochim. Biophys. Acta. 2017 1859:1900–1910. [PubMed]
2. Alpert E., Gruzman A., Totary H., Kaiser N., Reich R., Sasson S. A natural protective mechanism against hyperglycaemia in vascular endothelial and smooth-muscle cells: role of glucose and 12-hydroxyeicosatetraenoic acid. Biochem. J. 2002;362:413–422. [PubMed]
3. Segel I.H. Wiley Interscience Publication; New York, USA: 1975. Enzyme Kinetics, Behavior and Analysis of Rapid Equilibrium and Steady-state Enzyme Systems.
4. Cornish-Bowden A. A simple graphical method for determining the inhibition constants of mixed, uncompetitive and non-competitive inhibitors. Biochem. J. 1974;137:143–144. [PubMed]

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