Functional studies of threonine 310 mutations in Glut1

T310I is pathogenic, causing Glut1 deficiency.

Dong Wang, Juan M. Pascual, Pavel Iserovich, Hong Yang, L. Ma, Kunyan Kuang, Felipe A. Zuniga, Ruo Peng Sun, Kavita M. Swaroop, Jorge Fischbarg, Darryl C. De Vivo

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Abstract

We have previously reported on a patient with the Glut1 deficiency syndrome (Online Mendelian Inheritance in Man number 606777) carrying a heterozygous T310I missense mutation in the GLUT1 gene (Klepper, J., Wang, D., Fischbarg, J., Vera, J. C., Jarjour, I. T., O'Driscoll, K. R., and De Vivo, D. C. (1999) Neurochem. Res. 24, 587-594). To investigate the molecular basis for the associated functional deficit, we constructed T310A, T310S, and T310I human GLUT1 mutants for expression in Xenopus laevis oocytes via cRNA injection. For all mutants, glucose transport was decreased, and osmotic water permeability (Pf) was increased. Km values for 3-O-methylglucose (3-OMG) uptake under zero-trans influx and equilibrium exchange influx conditions were, respectively, 13 +/- 1 and 68 +/- 5 mm for wild-type Glut1, 5 +/- 1 and 25 +/- 6 mm for T310A, 6 +/- 3 and 30 +/- 6 mm for T310I, and 5 +/- 1 and 48 +/- 5 mm for T310S. Compared with wild-type Glut1, we determined the following. (a). Zero-trans and equilibrium exchange influx values of 3-OMG were significantly decreased, respectively, 15 and 5% in T310A, 8 and 3% in T310I, and 40 and 34% in T310S mutants. (b). Zero-trans efflux of 3-OMG and dehydroascorbic acid uptake were significantly decreased in mutants. (c). The relative Pf values for T310A, T310I, and T310S were increased 3-, 4.8-, and 3.5-fold compared with wild-type values. We found a very high negative correlation between the rate of glucose uptake and Pf (-0.93), and between hydropathy and uptake (-0.92), a moderate correlation between hydropathy and Pf (0.73), and a minimal correlation between uptake, Pf, and molecular weight. These findings are consistent with a central role for hydropathy rather than size at position 310 of this mutation.

Original languageEnglish (US)
Pages (from-to)49015-49021
Number of pages7
JournalThe Journal of biological chemistry
Volume278
Issue number49
StatePublished - Dec 5 2003

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3-O-Methylglucose
Threonine
Mutation
Dehydroascorbic Acid
Genetic Databases
Glucose
Complementary RNA
Xenopus laevis
Missense Mutation
Oocytes
Permeability
Genes
Molecular Weight
Molecular weight
Injections
Water

ASJC Scopus subject areas

  • Biochemistry

Cite this

Wang, D., Pascual, J. M., Iserovich, P., Yang, H., Ma, L., Kuang, K., ... De Vivo, D. C. (2003). Functional studies of threonine 310 mutations in Glut1: T310I is pathogenic, causing Glut1 deficiency. The Journal of biological chemistry, 278(49), 49015-49021.

Functional studies of threonine 310 mutations in Glut1 : T310I is pathogenic, causing Glut1 deficiency. / Wang, Dong; Pascual, Juan M.; Iserovich, Pavel; Yang, Hong; Ma, L.; Kuang, Kunyan; Zuniga, Felipe A.; Sun, Ruo Peng; Swaroop, Kavita M.; Fischbarg, Jorge; De Vivo, Darryl C.

In: The Journal of biological chemistry, Vol. 278, No. 49, 05.12.2003, p. 49015-49021.

Research output: Contribution to journalArticle

Wang, D, Pascual, JM, Iserovich, P, Yang, H, Ma, L, Kuang, K, Zuniga, FA, Sun, RP, Swaroop, KM, Fischbarg, J & De Vivo, DC 2003, 'Functional studies of threonine 310 mutations in Glut1: T310I is pathogenic, causing Glut1 deficiency.', The Journal of biological chemistry, vol. 278, no. 49, pp. 49015-49021.
Wang, Dong ; Pascual, Juan M. ; Iserovich, Pavel ; Yang, Hong ; Ma, L. ; Kuang, Kunyan ; Zuniga, Felipe A. ; Sun, Ruo Peng ; Swaroop, Kavita M. ; Fischbarg, Jorge ; De Vivo, Darryl C. / Functional studies of threonine 310 mutations in Glut1 : T310I is pathogenic, causing Glut1 deficiency. In: The Journal of biological chemistry. 2003 ; Vol. 278, No. 49. pp. 49015-49021.
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abstract = "We have previously reported on a patient with the Glut1 deficiency syndrome (Online Mendelian Inheritance in Man number 606777) carrying a heterozygous T310I missense mutation in the GLUT1 gene (Klepper, J., Wang, D., Fischbarg, J., Vera, J. C., Jarjour, I. T., O'Driscoll, K. R., and De Vivo, D. C. (1999) Neurochem. Res. 24, 587-594). To investigate the molecular basis for the associated functional deficit, we constructed T310A, T310S, and T310I human GLUT1 mutants for expression in Xenopus laevis oocytes via cRNA injection. For all mutants, glucose transport was decreased, and osmotic water permeability (Pf) was increased. Km values for 3-O-methylglucose (3-OMG) uptake under zero-trans influx and equilibrium exchange influx conditions were, respectively, 13 +/- 1 and 68 +/- 5 mm for wild-type Glut1, 5 +/- 1 and 25 +/- 6 mm for T310A, 6 +/- 3 and 30 +/- 6 mm for T310I, and 5 +/- 1 and 48 +/- 5 mm for T310S. Compared with wild-type Glut1, we determined the following. (a). Zero-trans and equilibrium exchange influx values of 3-OMG were significantly decreased, respectively, 15 and 5{\%} in T310A, 8 and 3{\%} in T310I, and 40 and 34{\%} in T310S mutants. (b). Zero-trans efflux of 3-OMG and dehydroascorbic acid uptake were significantly decreased in mutants. (c). The relative Pf values for T310A, T310I, and T310S were increased 3-, 4.8-, and 3.5-fold compared with wild-type values. We found a very high negative correlation between the rate of glucose uptake and Pf (-0.93), and between hydropathy and uptake (-0.92), a moderate correlation between hydropathy and Pf (0.73), and a minimal correlation between uptake, Pf, and molecular weight. These findings are consistent with a central role for hydropathy rather than size at position 310 of this mutation.",
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T2 - T310I is pathogenic, causing Glut1 deficiency.

AU - Wang, Dong

AU - Pascual, Juan M.

AU - Iserovich, Pavel

AU - Yang, Hong

AU - Ma, L.

AU - Kuang, Kunyan

AU - Zuniga, Felipe A.

AU - Sun, Ruo Peng

AU - Swaroop, Kavita M.

AU - Fischbarg, Jorge

AU - De Vivo, Darryl C.

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