Tryptophan scanning reveals dense packing of connexin transmembrane domains in gap junction channels composed of connexin32

Matthew J. Brennan, Jennifer Karcz, Nicholas R. Vaughn, Yvonne Woolwine-Cunningham, Adam D. DePriest, Yerko Escalona, Tomas Perez-Acle, I. Martha Skerrett

Resultado de la investigación: Contribución a una revistaArtículo

2 Citas (Scopus)

Resumen

Tryptophan was substituted for residues in all four transmembrane domains of connexin32. Function was assayed using dual cell two-electrode voltage clamp after expression in Xenopus oocytes. Tryptophan substitution was poorly tolerated in all domains, with the greatest impact in TM1 and TM4. For instance, in TM1, 15 substitutions were made, six abolished coupling and five others significantly reduced function. Only TM2 and TM3 included a distinct helical face that lacked sensitivity to tryptophan substitution. Results were visualized on a comparative model of Cx32 hemichannel. In this model, a region midway through the membrane appears highly sensitive to tryptophan substitution and includes residues Arg-32, Ile-33, Met-34, and Val-35. In the modeled channel, porefacing regions of TM1 and TM2 were highly sensitive to tryptophan substitution, whereas the lipid-facing regions of TM3 and TM4 were variably tolerant. Residues facing a putative intracellular water pocket (the IC pocket) were also highly sensitive to tryptophan substitution. Although future studies will be required to separate trafficking-defective mutants from those that alter channel function, a subset of interactions important for voltage gating was identified. Interactions important for voltage gating occurred mainly in the mid-region of the channel and focused on TM1. To determine whether results could be extrapolated to other connexins, TM1 of Cx43 was scanned revealing similar but not identical sensitivity to TM1 of Cx32.

Idioma originalInglés
Páginas (desde-hasta)17074-17084
Número de páginas11
PublicaciónJournal of Biological Chemistry
Volumen290
N.º28
DOI
EstadoPublicada - 10 jul 2015

Áreas temáticas de ASJC Scopus

  • Bioquímica
  • Medicina (todo)
  • Biología molecular
  • Biología celular

Huella Profundice en los temas de investigación de 'Tryptophan scanning reveals dense packing of connexin transmembrane domains in gap junction channels composed of connexin32'. En conjunto forman una huella única.

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    Brennan, M. J., Karcz, J., Vaughn, N. R., Woolwine-Cunningham, Y., DePriest, A. D., Escalona, Y., Perez-Acle, T., & Skerrett, I. M. (2015). Tryptophan scanning reveals dense packing of connexin transmembrane domains in gap junction channels composed of connexin32. Journal of Biological Chemistry, 290(28), 17074-17084. https://doi.org/10.1074/jbc.M115.650747