Structured Review

Stratagene y123w c1bα mutant
Distribution of open and closed loop conformations in <t>C1Bα.</t> (a) Histogram of the distances between the loop tips for the wt (yellow bars) and <t>Y123W</t> C1Bα (empty bars) observed during the interval between 2 and 10 ns of the MD trajectories. The distance between the loop tips is measured every 200 fs. Wt C1Bα shows a broad bimodal distribution centered at 12.5 Å and 9.5 Å, respectively. Y123W C1Bα has two preferred conformations: the open and closed, which are centered at 12.5 Å and 5 Å. (b) Histogram of the distances between the loop tips for the Y123W C1Bα (empty bars) observed during the trajectory interval between 10 and 18 ns, after the opening of the binding loops. The distance between the loop tips is measured every 200 fs. For comparison, the histogram generated using the three original 8 ns long trajectories of wt C1Bα is shown on the same plot (yellow bars). Both wt and Y123W C1Bα sample open or partially open conformations that show a bimodal distribution. Frequent transitions between open and partially open conformations are observed along the trajectories. Snapshots of the structures with closed and open loop conformations are shown in (c) and (d), respectively. Loop regions are highlighted in purple.
Y123w C1bα Mutant, supplied by Stratagene, used in various techniques. Bioz Stars score: 85/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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y123w c1bα mutant - by Bioz Stars, 2020-09
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1) Product Images from "Probing the Determinants of Diacylglycerol Binding Affinity in C1B domain of Protein Kinase C?"

Article Title: Probing the Determinants of Diacylglycerol Binding Affinity in C1B domain of Protein Kinase C?

Journal: Journal of molecular biology

doi: 10.1016/j.jmb.2011.03.020

Distribution of open and closed loop conformations in C1Bα. (a) Histogram of the distances between the loop tips for the wt (yellow bars) and Y123W C1Bα (empty bars) observed during the interval between 2 and 10 ns of the MD trajectories. The distance between the loop tips is measured every 200 fs. Wt C1Bα shows a broad bimodal distribution centered at 12.5 Å and 9.5 Å, respectively. Y123W C1Bα has two preferred conformations: the open and closed, which are centered at 12.5 Å and 5 Å. (b) Histogram of the distances between the loop tips for the Y123W C1Bα (empty bars) observed during the trajectory interval between 10 and 18 ns, after the opening of the binding loops. The distance between the loop tips is measured every 200 fs. For comparison, the histogram generated using the three original 8 ns long trajectories of wt C1Bα is shown on the same plot (yellow bars). Both wt and Y123W C1Bα sample open or partially open conformations that show a bimodal distribution. Frequent transitions between open and partially open conformations are observed along the trajectories. Snapshots of the structures with closed and open loop conformations are shown in (c) and (d), respectively. Loop regions are highlighted in purple.
Figure Legend Snippet: Distribution of open and closed loop conformations in C1Bα. (a) Histogram of the distances between the loop tips for the wt (yellow bars) and Y123W C1Bα (empty bars) observed during the interval between 2 and 10 ns of the MD trajectories. The distance between the loop tips is measured every 200 fs. Wt C1Bα shows a broad bimodal distribution centered at 12.5 Å and 9.5 Å, respectively. Y123W C1Bα has two preferred conformations: the open and closed, which are centered at 12.5 Å and 5 Å. (b) Histogram of the distances between the loop tips for the Y123W C1Bα (empty bars) observed during the trajectory interval between 10 and 18 ns, after the opening of the binding loops. The distance between the loop tips is measured every 200 fs. For comparison, the histogram generated using the three original 8 ns long trajectories of wt C1Bα is shown on the same plot (yellow bars). Both wt and Y123W C1Bα sample open or partially open conformations that show a bimodal distribution. Frequent transitions between open and partially open conformations are observed along the trajectories. Snapshots of the structures with closed and open loop conformations are shown in (c) and (d), respectively. Loop regions are highlighted in purple.

Techniques Used: Binding Assay, Generated

NMR-detected titration of the wt (a) and Y123W C1Bα (b) with DOG in the presence of DPC/DPS micelles. The binding process is intermediate-to-fast and slow-to-intermediate on the chemical-shift timescale for the wt and Y123W C1Bα, respectively. The insets show the differences in the binding regimes for the wt and mutant proteins using Leu122 as an example. Large chemical shift perturbations are observed in the ligand-bound versus apo-spectra.
Figure Legend Snippet: NMR-detected titration of the wt (a) and Y123W C1Bα (b) with DOG in the presence of DPC/DPS micelles. The binding process is intermediate-to-fast and slow-to-intermediate on the chemical-shift timescale for the wt and Y123W C1Bα, respectively. The insets show the differences in the binding regimes for the wt and mutant proteins using Leu122 as an example. Large chemical shift perturbations are observed in the ligand-bound versus apo-spectra.

Techniques Used: Nuclear Magnetic Resonance, Titration, Binding Assay, Mutagenesis

DOG binding curves for the wt and Y123W C1Bα detected by NMR and fluorescence spectroscopy. In (a) and (b), the absolute values of the 1 H and 15 N chemical shift changes, Δ 1 H and Δ 15 using the dissociation constant K d produced P 0 of 0.23 ± 0.07 μM and K d of 6.7 ± 16.4 nM. Large errors in K d indicate that, in this protein concentration range, the binding is still tight, and we can only put an upper limit of 0.23 μM onto the K d value.
Figure Legend Snippet: DOG binding curves for the wt and Y123W C1Bα detected by NMR and fluorescence spectroscopy. In (a) and (b), the absolute values of the 1 H and 15 N chemical shift changes, Δ 1 H and Δ 15 using the dissociation constant K d produced P 0 of 0.23 ± 0.07 μM and K d of 6.7 ± 16.4 nM. Large errors in K d indicate that, in this protein concentration range, the binding is still tight, and we can only put an upper limit of 0.23 μM onto the K d value.

Techniques Used: Binding Assay, Nuclear Magnetic Resonance, Fluorescence, Spectroscopy, Produced, Protein Concentration

Assessment of structural differences between wt and Y123W C1Bα using chemical shift perturbation analysis and RDCs. Structural Zn 2+ ions are shown as black spheres in (a), (c), and (d). Prolines and residues that are missing from the 15 N- 1 The only significant perturbation is observed at the mutation site. (b) Comparison of the 1 D NH
Figure Legend Snippet: Assessment of structural differences between wt and Y123W C1Bα using chemical shift perturbation analysis and RDCs. Structural Zn 2+ ions are shown as black spheres in (a), (c), and (d). Prolines and residues that are missing from the 15 N- 1 The only significant perturbation is observed at the mutation site. (b) Comparison of the 1 D NH

Techniques Used: Mutagenesis

Related Articles

Mutagenesis:

Article Title: Probing the Determinants of Diacylglycerol Binding Affinity in C1B domain of Protein Kinase C?
Article Snippet: .. Mutagenic DNA for the Y123W C1Bα mutant was constructed from the wt C1Bα gene using a Stratagene QuickChange™ site-directed mutagenesis kit and suitable PCR primers. .. Purified wt and mutagenic plasmids were transformed into BL21(DE3) E. coli cells.

Polymerase Chain Reaction:

Article Title: Probing the Determinants of Diacylglycerol Binding Affinity in C1B domain of Protein Kinase C?
Article Snippet: .. Mutagenic DNA for the Y123W C1Bα mutant was constructed from the wt C1Bα gene using a Stratagene QuickChange™ site-directed mutagenesis kit and suitable PCR primers. .. Purified wt and mutagenic plasmids were transformed into BL21(DE3) E. coli cells.

Construct:

Article Title: Probing the Determinants of Diacylglycerol Binding Affinity in C1B domain of Protein Kinase C?
Article Snippet: .. Mutagenic DNA for the Y123W C1Bα mutant was constructed from the wt C1Bα gene using a Stratagene QuickChange™ site-directed mutagenesis kit and suitable PCR primers. .. Purified wt and mutagenic plasmids were transformed into BL21(DE3) E. coli cells.

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    Stratagene y123w c1bα mutant
    Distribution of open and closed loop conformations in <t>C1Bα.</t> (a) Histogram of the distances between the loop tips for the wt (yellow bars) and <t>Y123W</t> C1Bα (empty bars) observed during the interval between 2 and 10 ns of the MD trajectories. The distance between the loop tips is measured every 200 fs. Wt C1Bα shows a broad bimodal distribution centered at 12.5 Å and 9.5 Å, respectively. Y123W C1Bα has two preferred conformations: the open and closed, which are centered at 12.5 Å and 5 Å. (b) Histogram of the distances between the loop tips for the Y123W C1Bα (empty bars) observed during the trajectory interval between 10 and 18 ns, after the opening of the binding loops. The distance between the loop tips is measured every 200 fs. For comparison, the histogram generated using the three original 8 ns long trajectories of wt C1Bα is shown on the same plot (yellow bars). Both wt and Y123W C1Bα sample open or partially open conformations that show a bimodal distribution. Frequent transitions between open and partially open conformations are observed along the trajectories. Snapshots of the structures with closed and open loop conformations are shown in (c) and (d), respectively. Loop regions are highlighted in purple.
    Y123w C1bα Mutant, supplied by Stratagene, used in various techniques. Bioz Stars score: 85/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/y123w c1bα mutant/product/Stratagene
    Average 85 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    y123w c1bα mutant - by Bioz Stars, 2020-09
    85/100 stars
      Buy from Supplier

    95
    Stratagene quickchange site directed mutagenesis kit
    Distribution of open and closed loop conformations in <t>C1Bα.</t> (a) Histogram of the distances between the loop tips for the wt (yellow bars) and <t>Y123W</t> C1Bα (empty bars) observed during the interval between 2 and 10 ns of the MD trajectories. The distance between the loop tips is measured every 200 fs. Wt C1Bα shows a broad bimodal distribution centered at 12.5 Å and 9.5 Å, respectively. Y123W C1Bα has two preferred conformations: the open and closed, which are centered at 12.5 Å and 5 Å. (b) Histogram of the distances between the loop tips for the Y123W C1Bα (empty bars) observed during the trajectory interval between 10 and 18 ns, after the opening of the binding loops. The distance between the loop tips is measured every 200 fs. For comparison, the histogram generated using the three original 8 ns long trajectories of wt C1Bα is shown on the same plot (yellow bars). Both wt and Y123W C1Bα sample open or partially open conformations that show a bimodal distribution. Frequent transitions between open and partially open conformations are observed along the trajectories. Snapshots of the structures with closed and open loop conformations are shown in (c) and (d), respectively. Loop regions are highlighted in purple.
    Quickchange Site Directed Mutagenesis Kit, supplied by Stratagene, used in various techniques. Bioz Stars score: 95/100, based on 3815 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/quickchange site directed mutagenesis kit/product/Stratagene
    Average 95 stars, based on 3815 article reviews
    Price from $9.99 to $1999.99
    quickchange site directed mutagenesis kit - by Bioz Stars, 2020-09
    95/100 stars
      Buy from Supplier

    Image Search Results


    Distribution of open and closed loop conformations in C1Bα. (a) Histogram of the distances between the loop tips for the wt (yellow bars) and Y123W C1Bα (empty bars) observed during the interval between 2 and 10 ns of the MD trajectories. The distance between the loop tips is measured every 200 fs. Wt C1Bα shows a broad bimodal distribution centered at 12.5 Å and 9.5 Å, respectively. Y123W C1Bα has two preferred conformations: the open and closed, which are centered at 12.5 Å and 5 Å. (b) Histogram of the distances between the loop tips for the Y123W C1Bα (empty bars) observed during the trajectory interval between 10 and 18 ns, after the opening of the binding loops. The distance between the loop tips is measured every 200 fs. For comparison, the histogram generated using the three original 8 ns long trajectories of wt C1Bα is shown on the same plot (yellow bars). Both wt and Y123W C1Bα sample open or partially open conformations that show a bimodal distribution. Frequent transitions between open and partially open conformations are observed along the trajectories. Snapshots of the structures with closed and open loop conformations are shown in (c) and (d), respectively. Loop regions are highlighted in purple.

    Journal: Journal of molecular biology

    Article Title: Probing the Determinants of Diacylglycerol Binding Affinity in C1B domain of Protein Kinase C?

    doi: 10.1016/j.jmb.2011.03.020

    Figure Lengend Snippet: Distribution of open and closed loop conformations in C1Bα. (a) Histogram of the distances between the loop tips for the wt (yellow bars) and Y123W C1Bα (empty bars) observed during the interval between 2 and 10 ns of the MD trajectories. The distance between the loop tips is measured every 200 fs. Wt C1Bα shows a broad bimodal distribution centered at 12.5 Å and 9.5 Å, respectively. Y123W C1Bα has two preferred conformations: the open and closed, which are centered at 12.5 Å and 5 Å. (b) Histogram of the distances between the loop tips for the Y123W C1Bα (empty bars) observed during the trajectory interval between 10 and 18 ns, after the opening of the binding loops. The distance between the loop tips is measured every 200 fs. For comparison, the histogram generated using the three original 8 ns long trajectories of wt C1Bα is shown on the same plot (yellow bars). Both wt and Y123W C1Bα sample open or partially open conformations that show a bimodal distribution. Frequent transitions between open and partially open conformations are observed along the trajectories. Snapshots of the structures with closed and open loop conformations are shown in (c) and (d), respectively. Loop regions are highlighted in purple.

    Article Snippet: Mutagenic DNA for the Y123W C1Bα mutant was constructed from the wt C1Bα gene using a Stratagene QuickChange™ site-directed mutagenesis kit and suitable PCR primers.

    Techniques: Binding Assay, Generated

    NMR-detected titration of the wt (a) and Y123W C1Bα (b) with DOG in the presence of DPC/DPS micelles. The binding process is intermediate-to-fast and slow-to-intermediate on the chemical-shift timescale for the wt and Y123W C1Bα, respectively. The insets show the differences in the binding regimes for the wt and mutant proteins using Leu122 as an example. Large chemical shift perturbations are observed in the ligand-bound versus apo-spectra.

    Journal: Journal of molecular biology

    Article Title: Probing the Determinants of Diacylglycerol Binding Affinity in C1B domain of Protein Kinase C?

    doi: 10.1016/j.jmb.2011.03.020

    Figure Lengend Snippet: NMR-detected titration of the wt (a) and Y123W C1Bα (b) with DOG in the presence of DPC/DPS micelles. The binding process is intermediate-to-fast and slow-to-intermediate on the chemical-shift timescale for the wt and Y123W C1Bα, respectively. The insets show the differences in the binding regimes for the wt and mutant proteins using Leu122 as an example. Large chemical shift perturbations are observed in the ligand-bound versus apo-spectra.

    Article Snippet: Mutagenic DNA for the Y123W C1Bα mutant was constructed from the wt C1Bα gene using a Stratagene QuickChange™ site-directed mutagenesis kit and suitable PCR primers.

    Techniques: Nuclear Magnetic Resonance, Titration, Binding Assay, Mutagenesis

    DOG binding curves for the wt and Y123W C1Bα detected by NMR and fluorescence spectroscopy. In (a) and (b), the absolute values of the 1 H and 15 N chemical shift changes, Δ 1 H and Δ 15 using the dissociation constant K d produced P 0 of 0.23 ± 0.07 μM and K d of 6.7 ± 16.4 nM. Large errors in K d indicate that, in this protein concentration range, the binding is still tight, and we can only put an upper limit of 0.23 μM onto the K d value.

    Journal: Journal of molecular biology

    Article Title: Probing the Determinants of Diacylglycerol Binding Affinity in C1B domain of Protein Kinase C?

    doi: 10.1016/j.jmb.2011.03.020

    Figure Lengend Snippet: DOG binding curves for the wt and Y123W C1Bα detected by NMR and fluorescence spectroscopy. In (a) and (b), the absolute values of the 1 H and 15 N chemical shift changes, Δ 1 H and Δ 15 using the dissociation constant K d produced P 0 of 0.23 ± 0.07 μM and K d of 6.7 ± 16.4 nM. Large errors in K d indicate that, in this protein concentration range, the binding is still tight, and we can only put an upper limit of 0.23 μM onto the K d value.

    Article Snippet: Mutagenic DNA for the Y123W C1Bα mutant was constructed from the wt C1Bα gene using a Stratagene QuickChange™ site-directed mutagenesis kit and suitable PCR primers.

    Techniques: Binding Assay, Nuclear Magnetic Resonance, Fluorescence, Spectroscopy, Produced, Protein Concentration

    Assessment of structural differences between wt and Y123W C1Bα using chemical shift perturbation analysis and RDCs. Structural Zn 2+ ions are shown as black spheres in (a), (c), and (d). Prolines and residues that are missing from the 15 N- 1 The only significant perturbation is observed at the mutation site. (b) Comparison of the 1 D NH

    Journal: Journal of molecular biology

    Article Title: Probing the Determinants of Diacylglycerol Binding Affinity in C1B domain of Protein Kinase C?

    doi: 10.1016/j.jmb.2011.03.020

    Figure Lengend Snippet: Assessment of structural differences between wt and Y123W C1Bα using chemical shift perturbation analysis and RDCs. Structural Zn 2+ ions are shown as black spheres in (a), (c), and (d). Prolines and residues that are missing from the 15 N- 1 The only significant perturbation is observed at the mutation site. (b) Comparison of the 1 D NH

    Article Snippet: Mutagenic DNA for the Y123W C1Bα mutant was constructed from the wt C1Bα gene using a Stratagene QuickChange™ site-directed mutagenesis kit and suitable PCR primers.

    Techniques: Mutagenesis