Supplementary MaterialsFIG?S1

Supplementary MaterialsFIG?S1. contigs JFZJ010000052 and JFZJ010000142 in the JA643 genome. Brands of phototrophs are symbolized in dark and MR-1 (heterotroph) and Ha sido-1 (chemolithotroph) are symbolized in reddish colored. (B) PioA-like homologs contain bigger N terminus and inner HR. PioA-like (dark) and MtrA-like (reddish colored) decaheme homologs are proven. Internal HR locations are highlighted in Ethacridine lactate yellow. Rpal, mutant and WT TIE-1. Heme-stainable band at 120 kDa (lane 2) shifts to >170 kDa (lane 4) after heat treatment. Porins, after heating, form an excessive network of hydrogen bonding that Rabbit polyclonal to Estrogen Receptor 1 holds -strands together into the -barrel shape, resulting in an SDS-resistant structure responsible for the shift in SDS-PAGE. The heat modifiability of holo-PioACB complex indicates that PioB is in its folded -barrel state (N. Noinaj, A. J. Kuszak, and S. K. Buchanan, Methods Mol Biol 1329:51C56, 2015, https://doi.org/10.1007/978-1-4939-2871-2_4). Download FIG?S4, PDF file, 1.1 MB. Copyright ? 2019 Gupta et al. This content is distributed under the terms of the Creative Commons Attribution 4.0 International license. FIG?S3. Production and characterization of recombinant holo-PioACr. (A) Coomassie and heme staining and Western blots (WB) with PioA-specific antibodies of His-enriched PioA protein samples. PioA-1 and PioA-2 are replicates. (B) UV-visible light spectral analysis of the affinity-purified holo-PioACr under aerobic conditions. The spectra for holo-PioACr are common of values were determined by Students test (1 star, < 0.05; 2 stars, < 0.01; 3 stars, < 0.001). Download FIG?S5, PDF file, 0.8 MB. Copyright ? 2019 Gupta et al. This content is distributed under the terms of the Creative Commons Attribution 4.0 International license. TABLE?S1. Average maximum current uptake (nA cm?2) under continuous light in -BECs for different bacterial strains. Download Table?S1, DOCX file, 0.01 MB. Copyright ? 2019 Gupta et al. This content is distributed under the terms of the Creative Commons Attribution 4.0 International license. FIG?S6. Ethacridine lactate Mass spectrometry (MS) analysis of the >170 kDa heme-stainable bands from DSM 162 and JA643. The sequences of PioA and PioB homologs from (A) and (B) are shown. Highlighted peptides (yellow) are the peptides detected by MS. The Ethacridine lactate CXXCH motifs are shown in red. The first three residues of HR that contains a PioA proteolysis site between Val242 and Ala243 in TIE-1 are highlighted in green. Download FIG?S6, PDF file, 0.9 MB. Copyright ? 2019 Gupta et al. This content is distributed under the terms of the Creative Commons Attribution 4.0 International license. FIG?S7. RaptorX-generated models of PioAC and PioB. (A) Predicted model of PioAC (green) and dodecaheme cytochrome GSU1996 (PDB accession number 3OV0, best template used in the model preparation; brown) were overlaid in Chimera using default parameters. (B) Surface of the PioAC-model from 1st CXXCH to the 10th CXXCH. CXXCH motifs (1 to 10) are shown in red, and the rest of the protein is shown in green. (C and D) Predicted model of PioB showing dimensions of protein surface (C) and pore (D). The N-terminal region of PioB which forms a plug-like structure is not shown in this model for simplicity. These models support the hypothesis that PioAC can insert entirely through PioB and that the complex (E) can take up electrons from the extracellular electron donors across the OM. Download FIG?S7, PDF file, 2.7 MB. Copyright ? 2019 Gupta et al. This content is distributed under the terms of the Creative Commons Attribution 4.0 International license. TABLE?S2. Relevant strains, plasmids, and primers employed Ethacridine lactate in this study. Download Table?S2, DOCX file, 0.02 MB. Copyright ? 2019 Gupta et al. This content is distributed under the conditions of the Innovative Commons Attribution 4.0 International permit. Text message?S1. Supplemental strategies. Download Text message S1, DOCX document, 0.02.

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