Journal: Nature Communications

Article Title: Integrin activation by the lipid molecule 25-hydroxycholesterol induces a proinflammatory response

doi: 10.1038/s41467-019-09453-x

Figure Lengend Snippet: 25HC interacts with α5β1 and αvβ3 integrins. a , b Biotinylation reactions were performed in the presence of either 25HC or DMSO (control). Cell lysates collected from BMDMs ( a ) or RAW 264.7 macrophages ( b ) incubated with biotinylated 25HC (biotin-25HC) or DMSO at 4 °C were precipitated (PPT) with avidin–agarose and subsequently immuno-blotted (IB) with either α5 integrin ( a ) or αv integrin ( b ) antibody. Total lysates were also blotted with α5 integrin ( a ), αv integrin ( b ), and actin ( a , b ) antibodies. c , d Biotin-25HC and DMSO (control) were incubated (PPT; precipitated) with avidin–agarose beads. 25HC conjugated beads were then incubated either with purified α5β1 integrin ( c ) or αvβ3 integrin ( d ). The avidin–agarose bound complex was subsequently immuno-blotted with either α5 integrin ( c ) or αv integrin ( d ) antibody. e , f Protein G-agarose beads were conjugated with either control IgG or antibody (Ab) specific for α5 integrin ( e ) and αv integrin ( f ). The beads were then incubated with either purified α5β1 integrin ( e ) or αvβ3 integrin ( f ) protein. Control beads (control IgG + purified integrin protein) and integrin protein bound beads (integrin-specific antibody + purified integrin protein) was incubated with tritiated 3 H-25HC. Bead-bound radioactivity (count per minute; CPM) was measured by scintillation counter. g Cell lysate collected from RAW 264.7 macrophages incubated with biotin-25HC, biotinylated 27-hydroxycholesterol (biotin-27HC), biotinylated 4β-hydroxycholesterol (biotin-4βHC) or DMSO (control) at 4 °C was precipitated (PPT) with avidin–agarose and subsequently immuno-blotted with α5 integrin antibody. Total cell lysate was also blotted with α5 integrin and β-actin antibodies. h Surface plasmon resonance (SPR) analyses were performed by injecting increasing concentrations of 25HC (16 nM, 40 nM, 160 nM, 640 nM, 1.6 µM) into both the αvβ3 integrin-immobilized and control surface of the CM5 chips. Each background-corrected sensorgram is a representative of replicate runs. i Binding of radioactive 3 H-25HC to immobilized αvβ3 integrin protein in the presence or absence of purified chemokine domain of fractalkine protein was assessed as described in the “methods” section. Immunoblot images are representative from three independent experiments. The radioactive values (mean ± standard deviation) are representative from two independent experiments. * p ≤ 0.05 using a Student’s t -test

Article Snippet: Purified human αvβ3 integrin protein (Yo proteins AB, Huddinge, Sweden) was covalently immobilized on a flow cell of CM5 chip (carboxy-methylated dextran coated) in 10 mM sodium acetate buffer, pH 4.0, using EDC/NHS amine coupling chemistry at 25 °C.

Techniques: Incubation, Avidin-Biotin Assay, Purification, Radioactivity, SPR Assay, Binding Assay, Western Blot, Standard Deviation

Journal: Nature Communications

Article Title: Integrin activation by the lipid molecule 25-hydroxycholesterol induces a proinflammatory response

doi: 10.1038/s41467-019-09453-x

Figure Lengend Snippet: Modeling identifies specific interactions between αvβ3 integrin and 25HC. a Ectodomain structure of αvβ3 integrin containing ‘RGD’’-binding site (site I) (blue) and site-II (red) residues highlighted by a layer of solvent molecule of radius 1.4 Å. The site-II-bound 25HC molecule is presented as a surface model in green and Mn 2+ ions from metal-ion-dependent ligand-binding site (MIDAS) and adjacent to MIDAS (ADMIDAS) sites of βI domain, β-propeller domain and genu knee region are presented in cyan. b Potential interacting residues for molecular recognition of αvβ3 integrin by 25HC were identified in a docking simulation. H-bonds are highlighted in dashed lines in red. c , d Ser399 of β-propeller (red), Ser162 (blue) and Ala263 (black) of βI domain make stable H-bond interactions with 3- and 25-hydroxyl groups of 25HC, respectively. The H-bond distances ( c ) and H-bond angles ( d ) are plotted against MD simulation time (200 ns). e The root-mean-square deviations (RMSD) measured for specificity-determining loop (SDL), α1- and α7 helices during the entire 200 ns long simulation. The SDL region undergoes significant conformational change (RMSD of ~6 Å) upon 25HC binding. f The conformational change in the SDL is accompanied by disruption of H-bond interaction between Tyr122-Thr182 residues, as well as change in the β-propeller blade that interacts with SDL. Movement of the regions from the beginning 0 ns (blue) to the end 200 ns (red) is indicated by arrows. g Correlation matrices (as heat maps) indicating correlated (red) or anti-correlated motions (blue) of α- and β-subunits of αvβ3 integrin in unbound and 25HC-bound states. h , i The distance between the amide NH of Q120 of the β-propeller domain and backbone carbonyl oxygen of P169 of the SDL. Breakage of this electrostatic interaction in the 25HC-bound protein (blue) during the simulation, around ~30 ns simulation causes significant increase in the distance, which remain intact in the unbound protein (magenta) during the entire 200 ns simulation time

Article Snippet: Purified human αvβ3 integrin protein (Yo proteins AB, Huddinge, Sweden) was covalently immobilized on a flow cell of CM5 chip (carboxy-methylated dextran coated) in 10 mM sodium acetate buffer, pH 4.0, using EDC/NHS amine coupling chemistry at 25 °C.

Techniques: Binding Assay, Ligand Binding Assay

Journal: Nature Communications

Article Title: Integrin activation by the lipid molecule 25-hydroxycholesterol induces a proinflammatory response

doi: 10.1038/s41467-019-09453-x

Figure Lengend Snippet: 25HC activates αvβ3 integrin in epithelial cell focal adhesions and macrophage podosomes. a Human lung epithelial cells (A549 cells) or differentiated THP-1 macrophages were subjected to various treatments and immunostained as indicated. In untreated (UT) A549 and THP-1 cells (first row), αvβ3 integrin, immunostained using the antibody LM609, was present in paxillin-positive focal adhesions and podosomes, respectively. In contrast, active αvβ3 integrin staining, indicated using the antibody AP5 (second and third row), was found almost exclusively in cells treated for 2 h with 0.5 µM 25HC compared with DMSO-treated controls. Merged images show the co-localization of αvβ3 integrin (total or activated) with focal adhesions/podosomes. Bar, 10 µm. b Quantification of the number of focal adhesions (FAs) and AP5 + FAs in A549 cells (three experiments, >12 cells per experiment; n ≥ 30). c Quantification of the number of podosomes and AP5 + podosomes in THP-1 cells (three experiments, 25 cells per experiment; n ≥ 70). Two- and One-tailed Wilcoxon rank-sum tests were performed to evaluate differences between total and AP5 + FA/podosome counts, respectively. Graphs depict mean ± SEM. * p ≤ 0.05

Article Snippet: Purified human αvβ3 integrin protein (Yo proteins AB, Huddinge, Sweden) was covalently immobilized on a flow cell of CM5 chip (carboxy-methylated dextran coated) in 10 mM sodium acetate buffer, pH 4.0, using EDC/NHS amine coupling chemistry at 25 °C.

Techniques: Staining, One-tailed Test

Journal: Nature Communications

Article Title: Integrin activation by the lipid molecule 25-hydroxycholesterol induces a proinflammatory response

doi: 10.1038/s41467-019-09453-x

Figure Lengend Snippet: α5β1 and αvβ3 integrins regulate 25HC-mediated proinflammatory response. a , c – g α5β1 and αvβ3 integrin-deficient cells were treated with 50 µM 25HC for 8 h to evaluate NFκB activation and proinflammatory response. a TNF secretion from 25HC-treated NR-9456 macrophages in the presence of either control IgG or α5β1 integrin blocking antibody (Ab). b Western blot analyses of α5 integrin protein expression in THP-1 cells transfected with either control siRNA (con siRNA) or α5 integrin-specific siRNA (α5 siRNA). c RT-PCR analyses of TNF expression in 25HC-treated THP-1 cells transfected with either control siRNA (con siRNA) or α5 integrin-specific siRNA (α5 siRNA). d Western blot and densitometric analyses of phospho-IκB status in 25HC-treated wild-type (WT) and α5 integrin knockout (KO) HAP1 cells. e TNF secretion from 25HC-treated WT and β3 integrin-deficient (β3 +/− cells) BMDMs. f , g TNF ( f ) and IL-6 ( g ) secretion from 25HC-treated THP-1 macrophages in the presence of either control IgG or αvβ3 integrin blocking antibody (Ab). RT-PCR images are representative from two independent experiments. The ELISA values (mean ± standard deviation) are representative from two or three independent experiments ( n = 4). * p ≤ 0.05 using a Student’s t -test. The densitometric quantification values for phospho-IκB (p-IκB) immunoblot represent the ratio of phospho-IκB:actin and the fold-induction was calculated after normalizing with the control 0 min group. The densitometric values represent the mean ± standard deviation from three independent studies. * p ≤ 0.05 using a Student’s t -test

Article Snippet: Purified human αvβ3 integrin protein (Yo proteins AB, Huddinge, Sweden) was covalently immobilized on a flow cell of CM5 chip (carboxy-methylated dextran coated) in 10 mM sodium acetate buffer, pH 4.0, using EDC/NHS amine coupling chemistry at 25 °C.

Techniques: Activation Assay, Blocking Assay, Western Blot, Expressing, Transfection, Reverse Transcription Polymerase Chain Reaction, Knock-Out, Enzyme-linked Immunosorbent Assay, Standard Deviation

Journal: Nature Communications

Article Title: Integrin activation by the lipid molecule 25-hydroxycholesterol induces a proinflammatory response

doi: 10.1038/s41467-019-09453-x

Figure Lengend Snippet: The 25HC-integrin-FAK signaling network is required for optimal Nod2 response. a IL-6 in serum of mice injected (via intraperitoneal route) with 25HC (50 mg/kg; 4 h) ( n = 5). b , c , e , g – m Cells were treated with 25 µg/ml of MDP for 8 h and d , f mice were injected intraperitoneally with 20 mg/kg of MDP for 4 h. C25H expression, 25HC production, FAK activation, and proinflammatory response were assessed. b RT-PCR analyses of C25H expression in MDP-treated wild-type (WT) and C25H knockout (KO) BMDMs. c 25HC levels in the medium supernatant of MDP-treated BMDMs. d 25HC levels in the serum of WT mice treated with MDP ( n = 4). e IL-6 secretion from WT and C25H KO BMDMs treated with MDP. f IL-6 levels in the serum of WT and C25H KO mice treated with MDP ( n = 4). g Western blot and densitometric analyses of FAK activation (phospho-FAK, Tyr397) status in MDP-treated THP-1 macrophages. h IL-6 production from MDP-treated WT and FAK KO MEFs (mouse embryo fibroblasts). i IL-6 secretion from MDP-treated NR-9456 macrophages in the presence of either control IgG or α5β1 integrin blocking antibody (Ab). j RT-PCR analyses of TNF expression in MDP-treated THP-1 cells transfected with either control siRNA (con siRNA) or α5 integrin-specific siRNA (α5 siRNA). k TNF secretion from MDP-treated WT and β3 integrin-deficient (β3 +/− cells) BMDMs. l TNF secretion from MDP-treated THP-1 macrophages in the presence of either control IgG or αvβ3 integrin blocking antibody. m Western blot and densitometric analyses of FAK activation (phospho-FAK, Tyr925) status in MDP-treated WT and C25H KO BMDM. RT-PCR images are representative from two independent experiments. The ELISA values (mean ± standard deviation) are representative from two or three independent experiments ( n = 4). * p ≤ 0.05 using a Student’s t -test. The densitometric quantification values for phospho-FAK (p-FAK) immunoblot represent the ratio of phospho-FAK:actin and the fold-induction was calculated after normalizing with the control untreated (UT) group. The densitometric values represent the mean ± standard deviation from three independent studies. * p ≤ 0.05 using a Student’s t- test

Article Snippet: Purified human αvβ3 integrin protein (Yo proteins AB, Huddinge, Sweden) was covalently immobilized on a flow cell of CM5 chip (carboxy-methylated dextran coated) in 10 mM sodium acetate buffer, pH 4.0, using EDC/NHS amine coupling chemistry at 25 °C.

Techniques: Injection, Expressing, Activation Assay, Reverse Transcription Polymerase Chain Reaction, Knock-Out, Western Blot, Blocking Assay, Transfection, Enzyme-linked Immunosorbent Assay, Standard Deviation

Journal: Nature Communications

Article Title: Integrin activation by the lipid molecule 25-hydroxycholesterol induces a proinflammatory response

doi: 10.1038/s41467-019-09453-x

Figure Lengend Snippet: The 25HC-integrin-FAK signaling network regulates proinflammatory response during virus infection. a 25HC levels in the medium supernatant of BMDMs infected with human respiratory syncytial virus (RSV). b TNF production from wild-type (WT) and C25H knockout (KO) BMDMs infected with RSV. c Western blot and densitometric analyses of FAK activation (phospho-FAK, Tyr925) status in RSV-infected BMDMs. d Western blot and densitometric analyses of FAK activation (phospho-FAK, Tyr925) status in influenza A virus (IAV)-infected BMDMs. e IL-6 production from RSV-infected WT and FAK KO MEFs. f IL-6 production from IAV-infected WT and FAK KO MEFs. g TNF production from RSV-infected BMDMs in the presence of either control IgG or α5β1 integrin blocking antibody (Ab). h IL-6 production from IAV-infected BMDMs in the presence of either control IgG or α5β1 integrin blocking antibody. i TNF secretion from IAV-infected WT and β3 integrin-deficient (β3 +/− cells) BMDMs. j IL-6 secretion from IAV-infected THP-1 cells in the presence of either control IgG or αvβ3 integrin blocking antibody. k IL-6 in the lungs of mice injected (via intratracheal or I.T route) with 25HC (5 mg/kg; 6 h) in the presence of either IgG or β1 integrin blocking antibody (Ab) administered to the mice via I.T route ( n = 4). l IL-6 in the lungs of mice injected (via I.T route) with 25HC (5 mg/kg; 6 h) in the presence of either vehicle (control) or FAK inhibitor (PND-1186) administered to the mice via I.T route ( n = 4). The ELISA values (mean ± standard deviation) are representative from two or three independent experiments ( n = 4). * p ≤ 0.05 using a Student’s t -test. The densitometric quantification values for phospho-FAK (p-FAK) immunoblot represent the ratio of phospho-FAK:actin and the fold-induction was calculated after normalizing with the control mock-infected group. The densitometric values represent the mean ± standard deviation from three independent studies. * p ≤ 0.05 using a Student’s t -test

Article Snippet: Purified human αvβ3 integrin protein (Yo proteins AB, Huddinge, Sweden) was covalently immobilized on a flow cell of CM5 chip (carboxy-methylated dextran coated) in 10 mM sodium acetate buffer, pH 4.0, using EDC/NHS amine coupling chemistry at 25 °C.

Techniques: Infection, Knock-Out, Western Blot, Activation Assay, Blocking Assay, Injection, Enzyme-linked Immunosorbent Assay, Standard Deviation

Journal: Nature Communications

Article Title: Integrin activation by the lipid molecule 25-hydroxycholesterol induces a proinflammatory response

doi: 10.1038/s41467-019-09453-x

Figure Lengend Snippet: A schematic model showing regulation of proinflammatory response by 25HC-integrin-FAK signaling network. Nod2 activation and virus (RSV and IAV) infection triggers expression of C25H, which results in production of 25HC. Extracellular 25HC activates integrin-FAK-NFκB signaling. Thus, 25HC links PRR (Nod2) pathway with integrin-FAK-NFκB signaling to confer optimal proinflammatory response following Nod2 activation and virus infection. INTG, integrin; FAK, focal adhesion kinase

Article Snippet: Purified human αvβ3 integrin protein (Yo proteins AB, Huddinge, Sweden) was covalently immobilized on a flow cell of CM5 chip (carboxy-methylated dextran coated) in 10 mM sodium acetate buffer, pH 4.0, using EDC/NHS amine coupling chemistry at 25 °C.

Techniques: Activation Assay, Infection, Expressing

Journal: Pharmaceuticals

Article Title: Proteolytically Derived Endogenous Angioinhibitors Originating from the Extracellular Matrix

doi: 10.3390/ph4121551

Figure Lengend Snippet: Extracellular Matrix derived Endogenous angioinhibitors and their mode of action.

Article Snippet: Prothrombin Kringle-2 , Prothrombin , αVβ3 integrin , Inhibits VEGF, MMP-2 and 9 expression, affects EC growth..

Techniques: Derivative Assay, Inhibition, Activation Assay, Binding Assay, Expressing

Journal: Innate Immunity

Article Title: RNA-activated protein kinase differentially modulates innate immune response mediated by supraphysiological concentrations of thyroid hormone

doi: 10.1177/1753425920955214

Figure Lengend Snippet: T3 induces ISG expression using non-genomic mechanisms involving integrin αVβ3 binding and activation of PLC–PI3K–AKT pathways. (a) HEK293FT cells were transfected with 1.5 µg of empty vector or cTHRα plasmid and protein extracts were processed for Western blot using Abs to THRα/β. Non-specific (NS) band is shown for equal loading. (b). HEK293FT cells were transfected with 1.0 μg of TATADR4-Luc in the presence or absence of 1.5 µg of empty vector or cTHRα plasmid. After 24 h of transfection, cells were treated with 75 µM T3 and harvested 16 h later for luciferase activity measurement. ** P < 0.01. NS: not significant. (c) HEK293FT cell were transfected with 1.0 µg of empty vector or cTHRα plasmid for 24 h. Cells were then treated with DMSO or 75 µM T3 for 16 h in the presence or absence of Poly IC. (d) 50 ng of IFIT-Promoter-Luc or 100 ng ISRE-Luc plasmids were transfected in HEK293FT cells followed by treatment with Poly IC in the presence or absence of DMSO or 75 µM T3. Cells were harvested and luciferase activity was measured. ** P < 0.01. (e) HEK293FT cells were treated with T3 (75 µM) or Poly IC either alone or in combination with each other in the presence of either DMSO or LY294002 (5 µM), U73122 (10 µM), or U0126 (5 µM) for 16 h. (f) HEK293FT cells were treated with T3 (75 µM) for indicated time periods. Proteins extracts were analyzed by Western blot using Abs to total AKT and phospho-AKT. (g) HEK293FT cells were treated for 16 h with DMSO or 75 µM T3 in the presence or absence of Poly IC and 50 µg/ml RGD peptide or a control peptide. RNA was isolated from the cell pellets (c, e, and g) and the expression of IFIT3 quantitated by real-time RT-PCR as described in the Methods section. All the data presented are in comparison to DMSO control groups. ** P < 0.01. NS: not significant.

Article Snippet: Cyclo (RGDfC) αVβ3 integrin-binding peptide (RGD) and Cyclo (-RADfK-), RGD negative control peptide, were obtained from Anaspec (Fermont, CA).

Techniques: Expressing, Binding Assay, Activation Assay, Transfection, Plasmid Preparation, Western Blot, Luciferase, Activity Assay, Control, Isolation, Quantitative RT-PCR, Comparison

Journal: Cancers

Article Title: Are Integrins Still Practicable Targets for Anti-Cancer Therapy?

doi: 10.3390/cancers11070978

Figure Lengend Snippet: Selected clinical trials of agents targeting integrins. Non-exhaustive listing of the recent most important clinical studies with integrin inhibitors and their salient features and results).

Article Snippet: Cilengitide (EMD 121974, anti-αVβ3/αVβ5 integrin cyclic peptide) (Merck-Serono): total no. of trials: 21 (+ 8 terminated) , , , , , , .

Techniques: Activity Assay

Journal: The Journal of Biological Chemistry

Article Title: Unique Disulfide Bonds in Epidermal Growth Factor (EGF) Domains of β3 Affect Structure and Function of αIIbβ3 and αvβ3 Integrins in Different Manner

doi: 10.1074/jbc.M111.311043

Figure Lengend Snippet: Surface expression of normal or mutant αIIbβ3 and αvβ3 integrins. Surface expression of αIIbβ3 or αvβ3 in BHK cells expressing normal or mutated β3 together with normal αIIb or αv were measured by flow cytometry using FITC-conjugated P2 antibody against αIIbβ3 or FITC-conjugated 23C6 antibody against αvβ3. The disulfide bonds that were disrupted by the mutations are depicted above the bars. The EGF domains that contain the disulfide bonds are shown below the bars. Error bars represent the mean ± S.E. of at least three experiments. Statistically significant differences were calculated by a two-tailed, unpaired t test. Asterisks denote a p value of <0.05.

Article Snippet: Molecular dynamics simulations of αIIbβ3 and αvβ3 protein fragments.

Techniques: Expressing, Mutagenesis, Flow Cytometry, Two Tailed Test

Journal: The Journal of Biological Chemistry

Article Title: Unique Disulfide Bonds in Epidermal Growth Factor (EGF) Domains of β3 Affect Structure and Function of αIIbβ3 and αvβ3 Integrins in Different Manner

doi: 10.1074/jbc.M111.311043

Figure Lengend Snippet: Activation state of normal or mutant αIIbβ3 and αvβ3 in BHK cells. Soluble fibrinogen binding was assessed by flow cytometry using FITC-conjugated anti-human fibrinogen as a secondary antibody in cells expressing either normal or mutated αIIbβ3 or αvβ3. The mutations depicted below the panels disrupted the disulfide bonds in the four EGF domains shown above each panel. Fibrinogen binding is presented as percent of αIIbβ3 or αvβ3 expression measured by P2 or 23C6 antibodies, respectively. Error bars represent mean ± S.E. of at least three experiments. Statistical significance was calculated by a two-tailed, unpaired t test (*, p < 0.05; **, p < 0.01; ***, p < 0.001).

Article Snippet: Molecular dynamics simulations of αIIbβ3 and αvβ3 protein fragments.

Techniques: Activation Assay, Mutagenesis, Binding Assay, Flow Cytometry, Expressing, Two Tailed Test

Journal: The Journal of Biological Chemistry

Article Title: Unique Disulfide Bonds in Epidermal Growth Factor (EGF) Domains of β3 Affect Structure and Function of αIIbβ3 and αvβ3 Integrins in Different Manner

doi: 10.1074/jbc.M111.311043

Figure Lengend Snippet: Activation of normal or mutant αIIbβ3 and αvβ3 by anti-LIBS6 antibody and Mn2+. Soluble fibrinogen binding to cells expressing either normal or mutated αIIbβ3 (A) or αvβ3 (B) was assessed by flow cytometry using FITC-conjugated anti-human fibrinogen as a secondary antibody with or without anti-LIBS6 and Mn2+. Fibrinogen binding is presented as the percent of αIIbβ3 or αvβ3 expression measured by P2 or 23C6 antibodies, respectively. Error bars represent the mean ± S.E. of at least three experiments. C, the correlation between fold increase in binding fibrinogen with anti-LIBS6 and Mn2+ and fibrinogen binding under non-activating conditions is shown. The regression analysis was calculated using the excel software. Depicted in a dashed circle is the value for the C523S/C544S αvβ3 mutant outlying the regression line.

Article Snippet: Molecular dynamics simulations of αIIbβ3 and αvβ3 protein fragments.

Techniques: Activation Assay, Mutagenesis, Binding Assay, Expressing, Flow Cytometry, Software

Journal: The Journal of Biological Chemistry

Article Title: Unique Disulfide Bonds in Epidermal Growth Factor (EGF) Domains of β3 Affect Structure and Function of αIIbβ3 and αvβ3 Integrins in Different Manner

doi: 10.1074/jbc.M111.311043

Figure Lengend Snippet: Molecular dynamics simulations of αIIbβ3 and αvβ3 protein fragments. A, shown is r.m.s.d. values (nm) of the backbone atoms from their initial coordinates as a function of simulation time. The r.m.s.d. of the mutated β3 fragments of αIIbβ3 (lower panel) and αvβ3 (upper panel) are shown. Notable was a significant conformational change in the mutated β3 fragment of αIIbβ3, whereas the mutated β3 fragment of αvβ3 returned to its original conformation. B, cluster distribution throughout the simulation was calculated with a r.m.s.d. cut-off value of 0.4 nm. It shows a stable dominant cluster in the αIIbβ3 mutated fragment (lower panel) and several clusters with none being dominant in the αvβ3 mutated fragment (upper panel). C, shown is the relative contribution of the different domains to the total protein r.m.s.d. The domains are color-coded. In αIIbβ3 (lower panel), the domains contribute differently to the conformational change, whereas in αvβ3 (upper panel) all domains changed to the same extent. D, superposition of mutant and WT most abundant conformations of β3 fragments of αIIbβ3 (lower panel) and αvβ3 (upper panel). The mutant fragments are shown in red, and the WT fragments are shown in blue. The focus is on the EGF-3 domain, which is presented in solid colors. The end of EGF-2 (left) and the beginning of EGF-4 (right) are presented in transparent colors. The WT Cys-523 and Cys-544 residues and the mutated Ser-523 and Ser-544 residues are indicated.

Article Snippet: Molecular dynamics simulations of αIIbβ3 and αvβ3 protein fragments.

Techniques: Mutagenesis

Journal: The Journal of Biological Chemistry

Article Title: Unique Disulfide Bonds in Epidermal Growth Factor (EGF) Domains of β3 Affect Structure and Function of αIIbβ3 and αvβ3 Integrins in Different Manner

doi: 10.1074/jbc.M111.311043

Figure Lengend Snippet: Classification of the four unique disulfide bonds by their configuration

Article Snippet: Molecular dynamics simulations of αIIbβ3 and αvβ3 protein fragments.

Techniques:

Journal: The Journal of Biological Chemistry

Article Title: Unique Disulfide Bonds in Epidermal Growth Factor (EGF) Domains of β3 Affect Structure and Function of αIIbβ3 and αvβ3 Integrins in Different Manner

doi: 10.1074/jbc.M111.311043

Figure Lengend Snippet: Effects of disulfide bond disruptions in the β3 subunit on αIIbβ3 and αvβ3 expression and activation ND, not determined.

Article Snippet: Molecular dynamics simulations of αIIbβ3 and αvβ3 protein fragments.

Techniques: Expressing, Activation Assay