The structural context of disease-causing mutations in gap junctions.
Gap junctions form intercellular channels that mediate metabolic and electrical signaling between neighboring cells in a tissue. Lack of an atomic resolution structure of the gap junction has made it difficult to identify interactions that stabilize its transmembrane domain. Using a recently computed model of this domain, which specifies the locations of each amino acid, we postulated the existence of several interactions and tested them experimentally. We introduced mutations within the transmembrane domain of the gap junction-forming protein connexin that were previously implicated in genetic diseases and that apparently destabilized the gap junction, as evidenced here by the absence of the protein from the sites of cell-cell apposition. The model structure helped identify positions on adjacent helices where second-site mutations restored membrane localization, revealing possible interactions between residue pairs. We thus identified two putative salt bridges and one pair involved in packing interactions in which one disease-causing mutation suppressed the effects of another. These results seem to reveal some of the physical forces that underlie the structural stability of the gap junction transmembrane domain and suggest that abrogation of such interactions bring about some of the effects of disease-causing mutations.
Fleishman
Sarel J
SJ
Department of Biochemistry, George S. Wise Faculty of Life Sciences, Sackler School of Medicine, Tel-Aviv University, 69978 Ramat Aviv, Israel.
Sabag
Adi D
AD
Ophir
Eran
E
Avraham
Karen B
KB
Ben-Tal
Nir
N
D002450:Cell Communication; D000072259:Connexin 26; D017630:Connexins / Q000235:genetics*; D017629:Gap Junctions; D006367:HeLa Cells; D006801:Humans; D008958:Models, Molecular; D008968:Molecular Conformation; D009154:Mutation; D011487:Protein Conformation; D017434:Protein Structure, Tertiary; D015398:Signal Transduction; D014162:Transfection; D000097002:Gap Junction beta-1 Protein
D000072259:Connexin 26; D017630:Connexins; D000097002:Gap Junction beta-1 Protein