Conductance of connexin hemichannels segregates with the first transmembrane segment.
Gap junction channels are intercellular channels that mediate the gated transfer of molecules between adjacent cells. To identify the domain determining channel conductance, the first transmembrane segment (M1) was reciprocally exchanged between Cx46 and Cx32E(1)43. The resulting chimeras exhibited conductances similar to that of the respective M1 donor. Furthermore, a chimera with the carboxy-terminal half of M1 in Cx46 replaced by that of Cx32 exhibited a conductance similar to that of Cx32E(1)43, whereas the chimera with only the amino-terminal half of M1 replaced retained the unitary conductance of wild-type Cx46. Extending the M1 domain swapping to other connexins by replacing the carboxy-terminal half of M1 in Cx46 with that of Cx37 yielded a chimera channel with increased unitary conductance close to that of Cx37. Furthermore, a point mutant of Cx46, with leucine substituted by glycine in position 35, displayed a conductance much larger than that of the wild type. Thus, the M1 segment, especially the second half, contains important determinants of conductance of the connexin channel.
Hu
Xinge
X
Department of Physiology and Biophysics, University of Miami, School of Medicine, Miami, Florida 33101, USA.
Ma
Meiyun
M
Dahl
Gerhard
G
D000595:Amino Acid Sequence; D000818:Animals; D001703:Biophysics / Q000379:methods; D017630:Connexins / Q000737:chemistry* / Q000235:genetics; D017629:Gap Junctions; D005998:Glycine / Q000737:chemistry; D015640:Ion Channel Gating; D007930:Leucine / Q000737:chemistry; D008564:Membrane Potentials; D008969:Molecular Sequence Data; D009154:Mutation; D009865:Oocytes / Q000378:metabolism; D018408:Patch-Clamp Techniques; D017434:Protein Structure, Tertiary; D012333:RNA, Messenger / Q000378:metabolism; D011993:Recombinant Fusion Proteins / Q000737:chemistry; D017422:Sequence Analysis, DNA; D017386:Sequence Homology, Amino Acid; D013096:Spermine / Q000494:pharmacology; D014981:Xenopus; D000097002:Gap Junction beta-1 Protein; D000097006:Gap Junction alpha-4 Protein
D017630:Connexins; D005998:Glycine; D007930:Leucine; D012333:RNA, Messenger; D011993:Recombinant Fusion Proteins; D013096:Spermine; D000097002:Gap Junction beta-1 Protein; D000097006:Gap Junction alpha-4 Protein; C082917:GJA3 protein, human