itc data analysis module origin 7.0  (OriginLab corp)

Journal: The Journal of Biological Chemistry

Article Title: The human RNA-binding protein and E3 ligase MEX-3C binds the MEX-3–recognition element (MRE) motif with high affinity

doi: 10.1074/jbc.M117.797746

Figure Lengend Snippet: The hMEX-3C KH domains bind the MRE10 RNA. A, sequence alignment of the KH domains of human MEX-3C and C. elegans MEX-3. Conserved residues are highlighted in gray. The GXXG motif is indicated by a red dashed box. The secondary structure elements of human MEX-3C KH domain assigned on the basis of the X-ray structure are shown at the top. B, the domain architecture of human MEX-3C and the sequence of MRE10 RNA. The black lines represent the hMEX-3C constructs used for ITC and structure determination. C, the ITC fitting result of hMEX-3C KH1/2 domains with MRE10 RNA. D, the ITC fitting results of individual hMEX-3C KH domain with MRE10a or MRE10b.

Article Snippet: Data analysis was done using the ITC data analysis module Origin 7.0 (MicroCal) provided by the manufacturer.

Techniques: Sequencing, Construct

Journal: The Journal of Biological Chemistry

Article Title: The human RNA-binding protein and E3 ligase MEX-3C binds the MEX-3–recognition element (MRE) motif with high affinity

doi: 10.1074/jbc.M117.797746

Figure Lengend Snippet: The key residues involved in RNA binding by MEX-3C KH domains. A, the ITC fitting curves of the wild-type hMEX-3C KH1/2 domains and its mutants to MRE10 RNA. B, the Kd value histogram of the wild-type KH1/2 domains and its mutants to MRE10. The error bars represent S.D.

Article Snippet: Data analysis was done using the ITC data analysis module Origin 7.0 (MicroCal) provided by the manufacturer.

Techniques: RNA Binding Assay

Journal: The Journal of Biological Chemistry

Article Title: The human RNA-binding protein and E3 ligase MEX-3C binds the MEX-3–recognition element (MRE) motif with high affinity

doi: 10.1074/jbc.M117.797746

Figure Lengend Snippet: Mutagenesis of MRE10 RNA. A, the MRE10 RNA was mutated systematically, and the Kd value for every single-nucleotide mutation was determined by ITC experiment. The change of standard free energy change is defined as ΔΔG0 = −RTln(1/Kd,mut) − [−RTln(1/Kd,wt)]. The bars represent the ΔΔG0 value, and the single-nucleotide mutants are indicated on the horizontal axis. The broken bars indicate the mutants that can hardly bind to hMEX-3C. B, upper, the RNA sequence of MRE10 and a series of spacing mutants with the background of cytosine are shown. The hMRE motif are shown in bold. Lower, the Kd value for each spacing mutant was determined by ITC experiment. The bars represent the ΔΔG0 value, and the spacing mutants are indicated on the horizontal axis. The error bars represent S.D.

Article Snippet: Data analysis was done using the ITC data analysis module Origin 7.0 (MicroCal) provided by the manufacturer.

Techniques: Mutagenesis, Sequencing