Gene (human) | KCNQ1 | HUGO Gene Nomenclature Committee (HGNC) | Gene ID: 3784; HGNC:6294 | |
Gene (human) | KCNE1 | HUGO Gene Nomenclature Committee (HGNC) | Gene ID: 3753; HGNC:6240 | |
Gene (human) | KCNE3 | HUGO Gene Nomenclature Committee (HGNC) | Gene ID: 10008; HGNC:6243 | |
Strain, strain background (Escherichia coli) | CT19 transaminase deficient strain | Dr. David Waugh, US National Cancer Institute PMID: 8914274 | | Used for special isotopic labeling of the KCNQ1 VSD for use in NMR studies. |
Strain, strain background (Escherichia coli) | Rosetta/ C43(DE3) | Sigma-Aldrich | Catalog number 70954 | Used for uniform isotopic labeling of KCNQ1 and amino acid-specific labeling for NMR. |
Recombinant DNA reagent | pET16b expression plasmid encoding tagged human KCNQ1- (100-249) | PMID: 24606221 | | Used to express the human KCNQ1 VSD for NMR structural studies |
Biological sample (include species here) | Xenopus oocytes (Xenopus laevis, female) | This paper | RRID:XEP_Xla | Xenopus laevis purchased from Nasco, Fort Atkinson, WI |
Recombinant DNA reagent | pcDNA3.1 encoding human KCNQ1 or KCNE1 | This paper | RRID:Addgene_111452 | For site-directed mutagenesis |
Commercial assay or kit | mMessage T7 polymerase kit | Applied Biosystems-Thermo Fisher Scientific | AM1344 | |
Chemical compound, drug | XE991 | Millipore Sigma | CAS #: 122955-42-4 | |
Chemical compound, drug | Chromanol 293B | Millipore Sigma | CAS #: 163163-23-3 | |
Chemical compound, drug | Alexa Fluor488 C5 maleimide | Molecular Probes, Eugene, OR | Catalog #: A10254 | |
Software | Topspin 3.2 | Bruker (Scientific Instruments Company) | RRID:SCR_014227 | NMR data collection and processing. |
Software | MDD and qMDD interface | URL: mddnmr.spektrino.com/ PMID: 21161328 | | NMR data processing |
Software | NMRFAM-SPARKY | PMID: 25505092 URL: https://nmrfam.wisc.edu/nmrfam-sparky-distribution/ | | NMR data analysis and resonance assignments. |
Software | TALOS-N | PMID:25502373 URL: spin.niddk.nih.gov/bax/software/TALOS-N/ | | Determination of secondary structure from NMR chemical shift data. |
Software | CHARMM-GUI | PMID: 25130509 URL: www.charmm-gui.org/ | | Preparation of starting structures of the KCNQ1 VSD in lipid bilayers for MD restrained MD simulations. |
Software | XPLOR-NIH | PMID: 28766807 URL: https://nmr.cit.nih.gov/xplor-nih/ | | Structure calculations using NMR data restraints |
Software | GPU-accelerated AMBER 16 | URL: https://ambermd.org/ PMID: 16200636 | | Program for execution of MD simulations. |
Software | Lipid 17 AMBER | PMID: 24803855 URL: https://ambermd.org/AmberModels.php | | Force field used for MD simulations. |
Software | CPPTRAJ | PMID: 26583988 URL:https://amber-md.github.io/ | | Analysis of MD trajectoriesfollowing simulations. |
Software | PatchMaster | HEKA | RRID:SCR_000034 | Electrophysiology data collection |
Software | IGOR | Wavemetrics, Lake Oswego, OR | RRID:SCR_000325 | Electrophysiology data analysis |
Software | Clampfit | Axon Instruments, Sunnyvale, CA | RRID:SCR_011323 | Electrophysiology data analysis |
Software | Sigmaplot | SPSS, San Jose, CA | RRID:SCR_003210 | Electrophysiology data analysis and visualization |
Software | MATLAB | MathWorks, MA | RRID:SCR_001622 | Electrophysiology data analysis |
Sequence-based reagent | For site-directed mutagenesis | This paper | PCR primers | PCR primers seq for mutations made in this study (each mutation utilizedtwo primers: b and c). E170R-b:cacgtacCTGgtcccgaagaacaccac; E170R-c: cgggacCAGgtacgtggtccgcctc; R237E-b: gcatcTCcaggatctgcaggaag; R237E-c:cagatcctgGAgatgctacacgtcgac F167R-b: ccgtcccgCGgaacaccaccagcac; F167R-c: gtgttcCGcgggacggagtacg Q234E-b:ggatctCcaggaagcggatgccc; Q234E-c: catccgcttcctgGagatcctgaggatgcta H240E-b: cggtcgacCTCtagcatcctcaggatc H240E-c: gctaGAGgtcgaccgccaggg D202N-b: cgatgaggtTAatgatggaaatgggcttc D202N-c: ccatcatTAacctcatcgtggtcgtg F351A-b: ggcaGCGcccgagccaagaatcc F351A-c: gctcgggCGCtgccctgaaggtgcag C214A-b: cttggaTcccacCGCgaggaccacca C214A-c: cGCGgtgggAtccaaggggcaggtg G219C-b: cctgaCacttggaTcccacCGC G219C-c: ggAtccaagtGtcaggtgtttgccacg C331A-b: gacagagaaTGCggaggcgatggtcttc C331A-c: ctccGCAttctctgtctttgccatc |