Structural signatures and membrane helix 4 in GLUT1

Inferences from human blood-brain glucose transport mutants

Juan M. Pascual, Dong Wang, Ru Yang, Lei Shi, Hong Yang, Darryl C. De Vivo

Research output: Contribution to journalArticle

31 Citations (Scopus)

Abstract

Exon IV of SLC2A1, a multiple facilitator superfamily (MFS) transporter gene, is particularly susceptible to mutations that cause GLUT1 deficiency syndrome, a human encephalopathy that results from decreased glucose flux through the blood-brain barrier. Genotyping of 100 patients revealed that in a third of them who harbor missense mutations in the GLUT1 transporter, transmembrane domain 4 (TM4), encoded by SLC2A1 exon IV, contains mutant residues that have the periodicity of one face of a kinked α-helix. Arg-126, located at the amino terminus of TM4, is the locus for most of the mutations followed by other arginine and glycine residues located elsewhere in the transporter but conserved among MFS proteins. The Arg-126 mutants were constructed and assayed for protein expression, targeting, and transport capacity in Xenopus oocytes. The role of charge at position 126, as well as its accessibility, was investigated in R126H by determining its activity as a function of extracellular pH. The results indicate that intracellular charges at the MFS TM2-3 and TM8-9 signature loops and flanking TMs 3, 5, and 6 are critical for the structure of GLUT1 as are TM glycines and that TM4, located at the catalytic core of MFS proteins, forms a helix that surfaces into the extracellular solution where another proton facilitates transport.

Original languageEnglish (US)
Pages (from-to)16732-16742
Number of pages11
JournalJournal of Biological Chemistry
Volume283
Issue number24
DOIs
StatePublished - Jun 13 2008

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Protein Transport
Glycine
Blood Glucose
Exons
Brain
Blood
Membranes
Glucose
Mutation
Brain Diseases
Periodicity
Missense Mutation
Xenopus
Blood-Brain Barrier
Oocytes
Arginine
Protons
Catalytic Domain
Proteins
Ports and harbors

ASJC Scopus subject areas

  • Biochemistry
  • Cell Biology
  • Molecular Biology

Cite this

Structural signatures and membrane helix 4 in GLUT1 : Inferences from human blood-brain glucose transport mutants. / Pascual, Juan M.; Wang, Dong; Yang, Ru; Shi, Lei; Yang, Hong; De Vivo, Darryl C.

In: Journal of Biological Chemistry, Vol. 283, No. 24, 13.06.2008, p. 16732-16742.

Research output: Contribution to journalArticle

Pascual, Juan M. ; Wang, Dong ; Yang, Ru ; Shi, Lei ; Yang, Hong ; De Vivo, Darryl C. / Structural signatures and membrane helix 4 in GLUT1 : Inferences from human blood-brain glucose transport mutants. In: Journal of Biological Chemistry. 2008 ; Vol. 283, No. 24. pp. 16732-16742.
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