human p85β ish2  (GenScript corporation)

Journal: Viruses

Article Title: Molecular Basis of the Ternary Interaction between NS1 of the 1918 Influenza A Virus, PI3K, and CRK

doi: 10.3390/v12030338

Figure Lengend Snippet: Interaction between 1918 NS1 and p85β (PDB ID: 6U28). ( A ) Crystal structure of the 1918 NS1 ED-CTT :p85β iSH2 complex. The position of residue 187 is shown as a blue sphere. Biolayer interferometry (BLI)-derived binding isotherms of ( B ) 1918 NS1E D-CTT and CRK-II, ( C ) 1918 NS1 ED-CTT and CRK-L. Insets: representative BLI sensorgrams with different analyte concentrations are shown by different colors. K D values and uncertainties are the global fitting result of three repeated data. ( D ) BLI sensorgram of 1918 NS1 ED-ΔCTT binding to CRK-L.

Article Snippet: Genes encoding 1918 NS1 ED-CTT (residues 86–230), human p85β iSH2 (residues 435–599), human CRK-II (residues 1–304) and CRK-L (residues 1–303) proteins were prepared by gene-synthesis service from Genscript (Piscataway, NJ, USA).

Techniques: Residue, Derivative Assay, Binding Assay

Journal: Viruses

Article Title: Molecular Basis of the Ternary Interaction between NS1 of the 1918 Influenza A Virus, PI3K, and CRK

doi: 10.3390/v12030338

Figure Lengend Snippet: Ternary interaction between 1918 NS1, p85β, and CRK. ( A ) Schematic showing the NS1-bridged (left) and p85β-bridged (right) ternary complex. BLI-derived binding isotherms between the 1918 NS1 ED-CTT :p85β complex and ( B ) CRK-II and ( C ) CRK-L. Insets: representative BLI sensorgrams with different analyte concentrations are shown by different colors. K D values and uncertainties are the global fitting result of three repeated data.

Article Snippet: Genes encoding 1918 NS1 ED-CTT (residues 86–230), human p85β iSH2 (residues 435–599), human CRK-II (residues 1–304) and CRK-L (residues 1–303) proteins were prepared by gene-synthesis service from Genscript (Piscataway, NJ, USA).

Techniques: Derivative Assay, Binding Assay

Journal: Viruses

Article Title: Molecular Basis of the Ternary Interaction between NS1 of the 1918 Influenza A Virus, PI3K, and CRK

doi: 10.3390/v12030338

Figure Lengend Snippet: The C-terminal tail (CTT) of 1918 NS1 mediates the interaction with CRK. ( A ) Crystal structure of the 1918 NS1 ED-CTT :p85β iSH2 complex. The region with missing electron densities are shown as a dashed black line. The PRM region is marked by a blue circle. ( B ) Representation of crystallographic B-factors of 1918 NS1 ED-CTT in complex with p85β. BLI-derived binding isotherms between p85β iSH2 and ( C ) 1918 NS1 ED-ΔCTT and ( D ) CRK-II. The result of p85β iSH2 and CRK-II was not fit because the affinity was too weak to fit reliably. Insets: representative BLI sensorgrams with different analyte concentrations shown by different colors. K D values and uncertainties are the global fitting result of three repeated data.

Article Snippet: Genes encoding 1918 NS1 ED-CTT (residues 86–230), human p85β iSH2 (residues 435–599), human CRK-II (residues 1–304) and CRK-L (residues 1–303) proteins were prepared by gene-synthesis service from Genscript (Piscataway, NJ, USA).

Techniques: Derivative Assay, Binding Assay

Journal: Viruses

Article Title: Molecular Basis of the Ternary Interaction between NS1 of the 1918 Influenza A Virus, PI3K, and CRK

doi: 10.3390/v12030338

Figure Lengend Snippet: Fuzzy electrostatic interaction between CRK and the 1918 NS1:p85β complex. ( A ) Crystal structure of the free nSH3 (left) of CRK-II and its complex (right) with PRM of 1918 NS1 (PDB ID: 5UL6). The protein surface is colored according to electric potential at neutral pH from -5 kT (red) to +5 kT (blue). (right panel) Positively charged residues of PRM are shown in blue. BLI-derived binding isotherms between the 1918 NS1 ED-CTT :p85β iSH2 complex and ( B ) CRK-II and ( C ) CRK-L in the presence of 1M NaCl. ( D ) BLI-derived binding isotherm between 1918 NS1 ED-CTT and p85β iSH2 in the presence of 1M NaCl. Insets: representative BLI sensorgrams with different analyte concentrations are shown by different colors. ( E ) A schematic model showing how 1918 NS1 might enhance activation of PI3K using hijacked CRK.

Article Snippet: Genes encoding 1918 NS1 ED-CTT (residues 86–230), human p85β iSH2 (residues 435–599), human CRK-II (residues 1–304) and CRK-L (residues 1–303) proteins were prepared by gene-synthesis service from Genscript (Piscataway, NJ, USA).

Techniques: Derivative Assay, Binding Assay, Activation Assay