Structural studies of N-terminal mutants of Connexin 32 using 1H NMR spectroscopy

B. D. Kalmatsky, Y. Batir, T. A. Bargiello, T. L. Dowd

Research output: Contribution to journalArticle

11 Citations (Scopus)

Abstract

The amino terminus of gap junction proteins, connexins, plays a fundamental role in voltage gating and ion permeation. We have previously shown with 1H NMR that the structure of the N-terminus of functional connexin molecules contains a flexible turn around G12 (Arch. Biochem. Biophys.490:9,2009) allowing the N-terminus to form a portion of the channel pore near the cytoplasmic entrance. The mutants of nonfunctional connexin molecules G12S and G12Y were found to prevent this turn. Previous functional studies of loci at which Cx32 mutations cause a peripheral neuropathy, Charcot-Marie-Tooth disease, have shown that G12S is not plasma membrane inserted. Presently, we solve the structure of nonfunctional Connexin 32 mutants W3D and Y7D which do not appear to be membrane inserted. Using 2D 1H NMR, we report that similar to G12S and G12Y, alterations in hydrophobic sidechain interactions disrupt (Y7D) or constrain (W3D) the flexible turn around G12. The alteration in the open turn around residue 12, observed in all nonfunctional mutants G12S, G12Y, W3D and Y7D correlates with loss of function. We propose that loss of the open turn causes the N-terminus to extend out of the channel pore and this misfolding may target mutants for destruction in the endoplasmic reticulum.

Original languageEnglish (US)
Pages (from-to)1-8
Number of pages8
JournalArchives of Biochemistry and Biophysics
Volume526
Issue number1
DOIs
StatePublished - Oct 1 2012

Fingerprint

Connexins
Nuclear magnetic resonance spectroscopy
Magnetic Resonance Spectroscopy
Nuclear magnetic resonance
Charcot-Marie-Tooth Disease
Molecules
Arches
Peripheral Nervous System Diseases
Cell membranes
Hydrophobic and Hydrophilic Interactions
Permeation
Endoplasmic Reticulum
Cell Membrane
Ions
Membranes
Mutation
connexin 32
Proton Magnetic Resonance Spectroscopy
Electric potential

Keywords

  • Atomic resolution structure
  • Connexins
  • Ion channels
  • NMR
  • Protein structure and function
  • Structure-function
  • Voltage dependent gating

ASJC Scopus subject areas

  • Biochemistry
  • Biophysics
  • Molecular Biology

Cite this

Structural studies of N-terminal mutants of Connexin 32 using 1H NMR spectroscopy. / Kalmatsky, B. D.; Batir, Y.; Bargiello, T. A.; Dowd, T. L.

In: Archives of Biochemistry and Biophysics, Vol. 526, No. 1, 01.10.2012, p. 1-8.

Research output: Contribution to journalArticle

Kalmatsky, B. D. ; Batir, Y. ; Bargiello, T. A. ; Dowd, T. L. / Structural studies of N-terminal mutants of Connexin 32 using 1H NMR spectroscopy. In: Archives of Biochemistry and Biophysics. 2012 ; Vol. 526, No. 1. pp. 1-8.
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