Heteromultimerization of prokaryotic bacterial cyclic nucleotide-gated (bCNG) ion channels, members of the mechanosensitive channel of small conductance (MscS) superfamily.

Traditionally, prokaryotic channels are thought to exist as homomultimeric assemblies, while many eukaryotic ion channels form complex heteromultimers. Here we demonstrate that bacterial cyclic nucleotide-gated channels likely form heteromultimers in vivo. Heteromultimer formation is indicated through channel modeling, pull-down assays, and real-time polymerase chain reaction analysis. Our observations demonstrate that prokaryotic ion channels can display complex behavior and regulation akin to that of their eukaryotic counterparts.

Ss-bCNGa: a unique member of the bacterial cyclic nucleotide gated (bCNG) channel family that gates in response to mechanical tension.

Bacterial cyclic nucleotide gated (bCNG) channels are generally a nonmechanosensitive subset of the mechanosensitive channel of small conductance (MscS) superfamily. bCNG channels are composed of an MscS channel domain, a linking domain, and a cyclic nucleotide binding domain. Among bCNG channels, the channel domain of Ss-bCNGa, a bCNG channel from Synechocystis sp. PCC 6803, is most identical to Escherichia coli (Ec) MscS. This channel also exhibits limited mechanosensation in response to osmotic downshock assays, making it the only known full-length bCNG channel to respond to hypoosmotic stress. Here, we compare and contrast the ability of Ss-bCNGa to gate in response to mechanical tension with Se-bCNG, a nonmechanosensitive bCNG channel, and Ec-MscS, a prototypical mechanosensitive channel. Compared with Ec-MscS, Ss-bCNGa only exhibits limited mechanosensation, which is most likely a result of the inability of Ss-bCNGa to form the strong lipid contacts needed for significant function. Unlike Ec-MscS, Ss-bCNGa displays a mechanical response that increases with protein expression level, which may result from channel clustering driven by interchannel cation-π interactions.

Defining the role of the tension sensor in the mechanosensitive channel of small conductance.

Mutations that alter the phenotypic behavior of the Escherichia coli mechanosensitive channel of small conductance (MscS) have been identified; however, most of these residues play critical roles in the transition between the closed and open states of the channel and are not directly involved in lipid interactions that transduce the tension response. In this study, we use molecular dynamic simulations to predict critical lipid interacting residues in the closed state of MscS. The physiological role of these residues was then investigated by performing osmotic downshock assays on MscS mutants where the lipid interacting residues were mutated to alanine. These experiments identified seven residues in the first and second transmembrane helices as lipid-sensing residues. The majority of these residues are hydrophobic amino acids located near the extracellular interface of the membrane. All of these residues interact strongly with the lipid bilayer in the closed state of MscS, but do not face the bilayer directly in structures associated with the open and desensitized states of the channel. Thus, the position of these residues relative to the lipid membrane appears related to the ability of the channel to sense tension in its different physiological states.

Angled-predrilling depth and mini-implant shape effects on the mechanical properties of self-drilling orthodontic mini-implants during the angled insertion procedure.

OBJECTIVE: To investigate the effects of orthodontic mini-implant (OMI) shape and angled-predrilling depth on the mechanical properties of OMIs during the angled insertion procedure. MATERIALS AND METHODS: A total of 30 OMIs (self-drilling type, 7 mm in length) were allocated into six groups according to the OMI shape (cylindrical or tapered) and angled-predrilling depth (control, 1.5-mm and 4.0-mm angled-predrilling; predrilled with 1-mm-diameter drill-bit), as follows: C-con, C-1.5, C-4.0, T-con, T-1.5, and T-4.0 groups (N  =  5 per group). The OMIs were installed at an angle of 60° in double-layer artificial bone blocks that simulated the cortical and cancellous bone (Sawbone(®)). Total insertion time (TIT), maximum insertion torque (MIT), total insertion energy (TIE), and inclination of the time-torque graph (INC) were measured. RESULTS: Within the same shape group, angled-predrilling had a shorter TIT than did the control (control vs 1.5; control vs 4.0; all P < .05). MIT and TIE decreased in the order of control, 1.5-mm, and 4.0-mm angled-predrilling (control vs 1.5; 1.5 vs 4.0; all P < .05), but INC increased from control to 1.5-mm angled-predrilling and decreased from 1.5-mm to 4.0-mm angled-predrilling within the same shape group (control vs 1.5, 1.5 vs 4.0; all P < .05). The MIT of the tapered group was greater than that of the cylindrical group (C-con vs T-con, C-1.5 vs T-1.5; all P < .05, C-4.0 vs T-4.0; P < .01). In the same angled-predrilling depth, no differences were observed in TIE between the cylindrical and tapered groups (C-1.5 vs T-1.5, C-4.0 vs T-4.0; all P > .05). CONCLUSIONS: In angled-predrilling insertion of OMIs into thick cortical bone, tapered OMIs might be a better choice than cylindrical OMIs for increasing primary stability because of higher MIT and similar TIE values.