Development of Cysteine-Free Fluorescent Proteins for the Oxidative Environment
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{"title"=>"Development of cysteine-free fluorescent proteins for the oxidative environment", "type"=>"journal", "authors"=>[{"first_name"=>"Takahisa", "last_name"=>"Suzuki", "scopus_author_id"=>"57199100740"}, {"first_name"=>"Seisuke", "last_name"=>"Arai", "scopus_author_id"=>"14520862000"}, {"first_name"=>"Mayumi", "last_name"=>"Takeuchi", "scopus_author_id"=>"36830761400"}, {"first_name"=>"Chiye", "last_name"=>"Sakurai", "scopus_author_id"=>"55226203400"}, {"first_name"=>"Hideaki", "last_name"=>"Ebana", "scopus_author_id"=>"55226451400"}, {"first_name"=>"Tsunehito", "last_name"=>"Higashi", "scopus_author_id"=>"8966067400"}, {"first_name"=>"Hitoshi", "last_name"=>"Hashimoto", "scopus_author_id"=>"55226327300"}, {"first_name"=>"Kiyotaka", "last_name"=>"Hatsuzawa", "scopus_author_id"=>"6701697718"}, {"first_name"=>"Ikuo", "last_name"=>"Wada", "scopus_author_id"=>"56548377300"}], "year"=>2012, "source"=>"PLoS ONE", "identifiers"=>{"pui"=>"364870702", "isbn"=>"1932-6203 (Electronic)\\r1932-6203 (Linking)", "issn"=>"19326203", "doi"=>"10.1371/journal.pone.0037551", "scopus"=>"2-s2.0-84861362282", "pmid"=>"22649538", "sgr"=>"84861362282"}, "id"=>"f83ddf85-f965-3e7c-baa7-65be23e7c78b", "abstract"=>"Molecular imaging employing fluorescent proteins has been widely used to highlight specific reactions or processes in various fields of the life sciences. Despite extensive improvements of the fluorescent tag, this technology is still limited in the study of molecular events in the extracellular milieu. This is partly due to the presence of cysteine in the fluorescent proteins. These proteins almost cotranslationally form disulfide bonded oligomers when expressed in the endoplasmic reticulum (ER). Although single molecule photobleaching analysis showed that these oligomers were not fluorescent, the fluorescent monomer form often showed aberrant behavior in folding and motion, particularly when fused to cysteine-containing cargo. Therefore we investigated whether it was possible to eliminate the cysteine without losing the brightness. By site-saturated mutagenesis, we found that the cysteine residues in fluorescent proteins could be replaced with specific alternatives while still retaining their brightness. cf(cysteine-free)SGFP2 showed significantly reduced restriction of free diffusion in the ER and marked improvement of maturation when fused to the prion protein. We further applied this approach to TagRFP family proteins and found a set of mutations that obtains the same level of brightness as the cysteine-containing proteins. The approach used in this study to generate new cysteine-free fluorescent tags should expand the application of molecular imaging to the extracellular milieu and facilitate its usage in medicine and biotechnology.", "link"=>"http://www.mendeley.com/research/development-cysteinefree-fluorescent-proteins-oxidative-environment", "reader_count"=>67, "reader_count_by_academic_status"=>{"Unspecified"=>1, "Professor > Associate Professor"=>3, "Librarian"=>1, "Researcher"=>15, "Student > Doctoral Student"=>5, "Student > Ph. D. Student"=>22, "Student > Postgraduate"=>1, "Other"=>1, "Student > Master"=>6, "Student > Bachelor"=>7, "Lecturer"=>1, "Professor"=>4}, "reader_count_by_user_role"=>{"Unspecified"=>1, "Professor > Associate Professor"=>3, "Librarian"=>1, "Researcher"=>15, "Student > Doctoral Student"=>5, "Student > Ph. D. Student"=>22, "Student > Postgraduate"=>1, "Other"=>1, "Student > Master"=>6, "Student > Bachelor"=>7, "Lecturer"=>1, "Professor"=>4}, "reader_count_by_subject_area"=>{"Engineering"=>1, "Unspecified"=>1, "Biochemistry, Genetics and Molecular Biology"=>16, "Agricultural and Biological Sciences"=>38, "Medicine and Dentistry"=>1, "Neuroscience"=>1, "Pharmacology, Toxicology and Pharmaceutical Science"=>1, "Chemical Engineering"=>1, "Chemistry"=>6, "Immunology and Microbiology"=>1}, "reader_count_by_subdiscipline"=>{"Engineering"=>{"Engineering"=>1}, "Medicine and Dentistry"=>{"Medicine and Dentistry"=>1}, "Neuroscience"=>{"Neuroscience"=>1}, "Chemistry"=>{"Chemistry"=>6}, "Immunology and Microbiology"=>{"Immunology and Microbiology"=>1}, "Agricultural and Biological Sciences"=>{"Agricultural and Biological Sciences"=>38}, "Biochemistry, Genetics and Molecular Biology"=>{"Biochemistry, Genetics and Molecular Biology"=>16}, "Unspecified"=>{"Unspecified"=>1}, "Pharmacology, Toxicology and Pharmaceutical Science"=>{"Pharmacology, Toxicology and Pharmaceutical Science"=>1}, "Chemical Engineering"=>{"Chemical Engineering"=>1}}, "reader_count_by_country"=>{"Netherlands"=>1, "Japan"=>1, "Poland"=>1, "Germany"=>1}, "group_count"=>2}

Scopus | Further Information

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Figshare

  • {"files"=>["https://ndownloader.figshare.com/files/634661"], "description"=>"<p>Brightness per molecule was estimated in the cytosol of living cells as described in <a href=\"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0037551#s4\" target=\"_blank\">Materials and Methods</a>, and plotted as box charts. Each fluorescent protein was expressed in a chamber stage for 3 hr at 37°C or for 3 hr at 28°C after the 3 hr expression period at 37°C. Twenty measurements were carried out for each protein. A box chart shows mean (cross), median (horizontal line), 25–75 (box) and 95 percentiles (bars) of molecular brightness.</p>", "links"=>[], "tags"=>["brightness", "cysteine-free", "fluorescent"], "article_id"=>305137, "categories"=>["Physics", "Biochemistry", "Biophysics", "Physiology", "Biotechnology", "Cell Biology"], "users"=>["Takahisa Suzuki", "Seisuke Arai", "Mayumi Takeuchi", "Chiye Sakurai", "Hideaki Ebana", "Tsunehito Higashi", "Hitoshi Hashimoto", "Kiyotaka Hatsuzawa", "Ikuo Wada"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0037551.g003", "stats"=>{"downloads"=>1, "page_views"=>17, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Molecular_brightness_of_cysteine_free_fluorescent_variants_/305137", "title"=>"Molecular brightness of cysteine-free fluorescent variants.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2012-05-23 01:25:37"}
  • {"files"=>["https://ndownloader.figshare.com/files/635544"], "description"=>"<p>Lysates of cells expressing ss-TagRFP, ss-cfTagRFP and ss-cgfTagRFP were subjected to immunoblot analysis using anti-TagRFP antibody (A). As with the GFP family proteins, ss-TagRFP showed extensive oligomers under non-reducing conditions (lane 2, <i>perpendicular line</i>). This is in contrast to ss-cfTagRFP (lanes 3 and 7) or ss-cgfTagRFP (lane 4 and 8) which showed identical patterns. The major band of ss-cgfTagRFP (<i>arrow</i>) migrated faster than ss-cfTagRFP. Tunicamycin treatment (5 µg/mL) for 8 hr caused faster migration (lane 10, “<i>aglyco</i>”) of ss-TagRFP (lane9, “<i>glycol</i>”). In contrast, ss-cgfTagRFP showed no difference in mobility (lanes 11 and 12). Bands of unknown origin that are inconsistently observed with ss-cgfTagRFP (lane 8) or ss-TagRFP (lanes 9 and 10) are indicated by an asterisk (*). Schematic drawings of mutations in cgfTagRFP are also shown. (B) Excitation and emission spectra of newly developed fluorescent proteins. Normalized absorption (<i>left</i>) or emission spectra (<i>right</i>) of the purified fluorescent proteins. (C) Localization of ss-cgfTagRFP in cells treated with brefeldin A. Fluorescence of ss-cgfTagRFP almost completely matched with anti-calnexin antibody immunofluorescence.</p>", "links"=>[], "tags"=>["cgftagrfp"], "article_id"=>306031, "categories"=>["Physics", "Biochemistry", "Biophysics", "Physiology", "Biotechnology", "Cell Biology"], "users"=>["Takahisa Suzuki", "Seisuke Arai", "Mayumi Takeuchi", "Chiye Sakurai", "Hideaki Ebana", "Tsunehito Higashi", "Hitoshi Hashimoto", "Kiyotaka Hatsuzawa", "Ikuo Wada"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0037551.g008", "stats"=>{"downloads"=>1, "page_views"=>8, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Development_of_cgfTagRFP_and_cgfmKate2_/306031", "title"=>"Development of cgfTagRFP and cgfmKate2.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2012-05-23 01:40:31"}
  • {"files"=>["https://ndownloader.figshare.com/files/635680"], "description"=>"a<p>Absolute quantum yield corrected with reabsorption and reemission (the excitation maximum in parentheses).</p>b<p>Extinction coefficient measured with the absorbance maximum.</p>c<p>Photostability. Time to bleach 1/e of fluorescence relative to EGFP (SGFP2, cfSGFP2) or TagRFP expressed in COS7 cells (cgfTagRFP, mKate2 and cgfmKate2).</p>d<p>Photon counts per second per molecule (cpsm) of fluorescent proteins in COS7 cells at 37°C (<a href=\"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0037551#pone-0037551-g003\" target=\"_blank\">Figure 3</a>).</p>e<p>Cited literature (<a href=\"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0037551#pone.0037551-Shcherbo2\" target=\"_blank\">[41]</a>).</p>", "links"=>[], "tags"=>["properties", "fluorescent"], "article_id"=>306160, "categories"=>["Physics", "Biochemistry", "Biophysics", "Physiology", "Biotechnology", "Cell Biology"], "users"=>["Takahisa Suzuki", "Seisuke Arai", "Mayumi Takeuchi", "Chiye Sakurai", "Hideaki Ebana", "Tsunehito Higashi", "Hitoshi Hashimoto", "Kiyotaka Hatsuzawa", "Ikuo Wada"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0037551.t001", "stats"=>{"downloads"=>1, "page_views"=>15, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Photo_chemical_properties_of_fluorescent_proteins_/306160", "title"=>"Photo-chemical properties of fluorescent proteins.", "pos_in_sequence"=>0, "defined_type"=>3, "published_date"=>"2012-05-23 01:42:40"}
  • {"files"=>["https://ndownloader.figshare.com/files/634378"], "description"=>"<p>Various GFP family proteins as indicated in the figure were targeted to the ER by a signal sequence (“ss”) and subjected to SDS-PAGE after SDS treatment at 70°C for 10 min with (lanes 1–8) or without (lanes 9–16) reduction as described in <a href=\"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0037551#s4\" target=\"_blank\">Materials and Methods</a>. Shown is an immunoblot probed with anti-GFP antibody. The position of each oligomer is indicated. Vehicle: mock-transfection. *: unidentified bands. The numbers to the left indicate the positions of molecular mass markers. When molecules contained two cysteine residues, they formed higher molecular mass oligomers (lanes 10, 11, 12 and 13) whereas molecules containing a single cysteine formed a dimer (lanes 14 and 16). The monomer form SGFP2(C48S/C70M) (lane 15) is named cfSGFP2.</p>", "links"=>[], "tags"=>["gfp", "proteins"], "article_id"=>304861, "categories"=>["Physics", "Biochemistry", "Biophysics", "Physiology", "Biotechnology", "Cell Biology"], "users"=>["Takahisa Suzuki", "Seisuke Arai", "Mayumi Takeuchi", "Chiye Sakurai", "Hideaki Ebana", "Tsunehito Higashi", "Hitoshi Hashimoto", "Kiyotaka Hatsuzawa", "Ikuo Wada"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0037551.g001", "stats"=>{"downloads"=>1, "page_views"=>16, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Oligomerization_of_GFP_family_proteins_in_the_ER_/304861", "title"=>"Oligomerization of GFP family proteins in the ER.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2012-05-23 01:21:01"}
  • {"files"=>["https://ndownloader.figshare.com/files/635303"], "description"=>"<p>COS7 cells expressing prion protein fused to either ss-cfSGFP2 (<i>left</i>) or ss-SGFP2 (<i>right</i>) were immunostained with GFP antibody and the epifluorescent (<i>red</i>) or autofluorescent (<i>green</i>) images were observed by confocal microscopy as described in <a href=\"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0037551#s4\" target=\"_blank\">Materials and Methods</a>. In ss-cfSGFP2-prion fusion protein, most of the GFP signal showed colocalization with the anti-GFP antibody signal (<i>merge, left</i>) with the exception of a perinuclear, Golgi-like region (see <i>Results</i>). In contrast, the ss-SGFP2-prion fusion protein showed markedly distinct images (<i>merge, right</i>). The confocal pinhole was set to 1.0 Airy Units.</p>", "links"=>[], "tags"=>["sgfp2-tagged"], "article_id"=>305788, "categories"=>["Physics", "Biochemistry", "Biophysics", "Physiology", "Biotechnology", "Cell Biology"], "users"=>["Takahisa Suzuki", "Seisuke Arai", "Mayumi Takeuchi", "Chiye Sakurai", "Hideaki Ebana", "Tsunehito Higashi", "Hitoshi Hashimoto", "Kiyotaka Hatsuzawa", "Ikuo Wada"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0037551.g007", "stats"=>{"downloads"=>0, "page_views"=>14, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Mistargeting_of_SGFP2_tagged_prion_/305788", "title"=>"Mistargeting of SGFP2-tagged prion.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2012-05-23 01:36:28"}
  • {"files"=>["https://ndownloader.figshare.com/files/634500"], "description"=>"<p>The indicated fluorescent proteins were expressed in cells and the cells were then lysed and analyzed by immunoblotting using either anti-GFP antibody (lanes 1–8) or anti-RFP antibody (lanes 9–16). As in <a href=\"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0037551#pone-0037551-g001\" target=\"_blank\">Figure 1</a>, samples were SDS-treated with (lanes 5–12) or without (lanes 1–4 and 13–16) reduction. “Cys:1″ indicates fluorescent proteins containing C48S (SGFP2) or S3C (mCherry) and SGFP2“Cys:2″ is the wild type SGFP2. In the lanes where two proteins are listed, both expression plasmids were cotransfected. Std: molecular mass markers as indicated in the figure. As neither antibody cross-reacted and indeed showed no reaction in lane 6, then the band in lane 2 is assumed to be a leaked sample from the adjacent lanes. mCherry(Cys:1) formed a disulfide bonded dimer (lane14). Since the molecular masses of SGFP2 and the mCherry oligomer are different (lane 1 or 5 versus lane 10 or 14), then heterooligomers consisting of SGFP2 and mCherry should be distinguishable from each homooligomer. As the band profiles of lane 1 (SGFP2(Cys:1) alone) and lane 3 (SGFP2(Cys:1)+mCherry(Cys:1)) were almost identical, then a disulfide bond was not formed between SGFP2(Cys:1) and mCherry(Cys:1). The same is true for lanes 14 (mCherry(Cys:1)) and 15 (SGFP2(Cys:1)+mCherry(Cys:1)) and lane 16 (SGFP2(Cys:2)+mCherry(Cys:1)).</p>", "links"=>[], "tags"=>["fluorescent", "proteins"], "article_id"=>304982, "categories"=>["Physics", "Biochemistry", "Biophysics", "Physiology", "Biotechnology", "Cell Biology"], "users"=>["Takahisa Suzuki", "Seisuke Arai", "Mayumi Takeuchi", "Chiye Sakurai", "Hideaki Ebana", "Tsunehito Higashi", "Hitoshi Hashimoto", "Kiyotaka Hatsuzawa", "Ikuo Wada"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0037551.g002", "stats"=>{"downloads"=>0, "page_views"=>16, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Oligomer_formation_of_fluorescent_proteins_is_specific_/304982", "title"=>"Oligomer formation of fluorescent proteins is specific.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2012-05-23 01:23:02"}
  • {"files"=>["https://ndownloader.figshare.com/files/634882"], "description"=>"<p>Cells expressing ss-SGFP2 were extensively fixed for 4 days with paraformaldehyde as described in <a href=\"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0037551#s4\" target=\"_blank\">Materials and Methods</a> and single molecules were directly visualized by total reflection fluorescence microscopy. (A) When 400 frames were recorded at a frame rate of 250 Hz, 4 spots disappeared from the first frame in the recorded 1.6 sec (indicated by arrows). The width and height of an image is 13.6 µm. (B) Kymograph of each spot (1–4) from panel A. A single pixel image of the Y axis is lined along progression of time. (C) Quantification of the fluorescent signal in panel B. All spots were photobleached in a single step.</p>", "links"=>[], "tags"=>["photobleaching", "sgfp2"], "article_id"=>305363, "categories"=>["Physics", "Biochemistry", "Biophysics", "Physiology", "Biotechnology", "Cell Biology"], "users"=>["Takahisa Suzuki", "Seisuke Arai", "Mayumi Takeuchi", "Chiye Sakurai", "Hideaki Ebana", "Tsunehito Higashi", "Hitoshi Hashimoto", "Kiyotaka Hatsuzawa", "Ikuo Wada"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0037551.g004", "stats"=>{"downloads"=>0, "page_views"=>1, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Single_step_photobleaching_of_SGFP2_in_the_ER_/305363", "title"=>"Single step photobleaching of SGFP2 in the ER.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2012-05-23 01:29:23"}
  • {"files"=>["https://ndownloader.figshare.com/files/327769", "https://ndownloader.figshare.com/files/327837", "https://ndownloader.figshare.com/files/327892", "https://ndownloader.figshare.com/files/327928", "https://ndownloader.figshare.com/files/327992"], "description"=>"<div><p>Molecular imaging employing fluorescent proteins has been widely used to highlight specific reactions or processes in various fields of the life sciences. Despite extensive improvements of the fluorescent tag, this technology is still limited in the study of molecular events in the extracellular milieu. This is partly due to the presence of cysteine in the fluorescent proteins. These proteins almost cotranslationally form disulfide bonded oligomers when expressed in the endoplasmic reticulum (ER). Although single molecule photobleaching analysis showed that these oligomers were not fluorescent, the fluorescent monomer form often showed aberrant behavior in folding and motion, particularly when fused to cysteine-containing cargo. Therefore we investigated whether it was possible to eliminate the cysteine without losing the brightness. By site-saturated mutagenesis, we found that the cysteine residues in fluorescent proteins could be replaced with specific alternatives while still retaining their brightness. cf(cysteine-free)SGFP2 showed significantly reduced restriction of free diffusion in the ER and marked improvement of maturation when fused to the prion protein. We further applied this approach to TagRFP family proteins and found a set of mutations that obtains the same level of brightness as the cysteine-containing proteins. The approach used in this study to generate new cysteine-free fluorescent tags should expand the application of molecular imaging to the extracellular milieu and facilitate its usage in medicine and biotechnology.</p> </div>", "links"=>[], "tags"=>["cysteine-free", "fluorescent", "proteins", "oxidative"], "article_id"=>124708, "categories"=>["Physics", "Biochemistry", "Biophysics", "Physiology", "Biotechnology", "Cell Biology"], "users"=>["Takahisa Suzuki", "Seisuke Arai", "Mayumi Takeuchi", "Chiye Sakurai", "Hideaki Ebana", "Tsunehito Higashi", "Hitoshi Hashimoto", "Kiyotaka Hatsuzawa", "Ikuo Wada"], "doi"=>["https://dx.doi.org/10.1371/journal.pone.0037551.s001", "https://dx.doi.org/10.1371/journal.pone.0037551.s002", "https://dx.doi.org/10.1371/journal.pone.0037551.s003", "https://dx.doi.org/10.1371/journal.pone.0037551.s004", "https://dx.doi.org/10.1371/journal.pone.0037551.s005"], "stats"=>{"downloads"=>7, "page_views"=>23, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/Development_of_Cysteine_Free_Fluorescent_Proteins_for_the_Oxidative_Environment/124708", "title"=>"Development of Cysteine-Free Fluorescent Proteins for the Oxidative Environment", "pos_in_sequence"=>0, "defined_type"=>4, "published_date"=>"2012-05-23 01:18:28"}
  • {"files"=>["https://ndownloader.figshare.com/files/634990"], "description"=>"<p>The autocorrelation function of SGFP (n = 146) or cfSGFP2 (n = 172) in the ER of living cells was measured as described in <a href=\"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0037551#s4\" target=\"_blank\">Materials and Methods</a>. Shown are the normalized mean (solid lines) and CI95 (perpendicular bars) of the measurements (A). Best fits and residuals for diffusion time <i>τ<sub>D</sub></i> and fraction were obtained when the three component diffusion model was applied (B). ss-cfSGFP2 showed a steeper decay compared to ss-SGFP2.</p>", "links"=>[], "tags"=>["restricts", "diffusion"], "article_id"=>305473, "categories"=>["Physics", "Biochemistry", "Biophysics", "Physiology", "Biotechnology", "Cell Biology"], "users"=>["Takahisa Suzuki", "Seisuke Arai", "Mayumi Takeuchi", "Chiye Sakurai", "Hideaki Ebana", "Tsunehito Higashi", "Hitoshi Hashimoto", "Kiyotaka Hatsuzawa", "Ikuo Wada"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0037551.g005", "stats"=>{"downloads"=>1, "page_views"=>9, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Cysteine_restricts_simple_diffusion_in_the_ER_/305473", "title"=>"Cysteine restricts simple diffusion in the ER.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2012-05-23 01:31:13"}
  • {"files"=>["https://ndownloader.figshare.com/files/635106"], "description"=>"<p>Lysates of cells expressing the prion fusion proteins indicated in the figure were analyzed by Western blotting as described in <a href=\"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0037551#s4\" target=\"_blank\">Materials and Methods</a>. (A) SDS-solubilized lysates were treated either without DTT (lanes1–4 and 9–12) or with DTT (lanes 5–8 and 13–16) at either 70°C or 95°C for 10 min. Prion protein fused to ss-cfSGFP2 migrated on the gel at the expected size of 58 kDa under all conditions (lanes 2, 6, 10 and 14). In contrast, prion protein fused to ss-SGFP2 or ss-SGFP2(C48S) showed intense fast migrating bands (lanes 3, 4, 7 and 8) when samples were treated at 70°C, but not at 95°C (lanes 11, 12, 15 and 16). * indicates an unknown form of the prion fusion protein which was not consistently observed. (B) Cells were incubated with or without 100 nM MG132 for 4 hr as indicated in the figure and, as in (A), cell lysates were treated under four different conditions. When MG132 was included, a set of faster migrating bands was not diminished completely by a 95°C treatment (lane13, <i>arrowhead</i>).</p>", "links"=>[], "tags"=>["aberrant", "sds-resistant", "sgfp2-prion", "fusion", "cfsgfp2-prion"], "article_id"=>305586, "categories"=>["Physics", "Biochemistry", "Biophysics", "Physiology", "Biotechnology", "Cell Biology"], "users"=>["Takahisa Suzuki", "Seisuke Arai", "Mayumi Takeuchi", "Chiye Sakurai", "Hideaki Ebana", "Tsunehito Higashi", "Hitoshi Hashimoto", "Kiyotaka Hatsuzawa", "Ikuo Wada"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0037551.g006", "stats"=>{"downloads"=>1, "page_views"=>12, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Formation_of_an_aberrant_SDS_resistant_structure_in_SGFP2_prion_fusion_protein_but_not_in_cfSGFP2_prion_fusion_protein_/305586", "title"=>"Formation of an aberrant SDS-resistant structure in SGFP2-prion fusion protein but not in cfSGFP2-prion fusion protein.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2012-05-23 01:33:06"}

PMC Usage Stats | Further Information

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Relative Metric

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