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Impaired RIPK2 binding of mutant proteins was not merely due to protein instability as demonstrated by similar expression levels (Figure 4, right panel).
Binding of mutant proteins was normalized to binding of MBP-P66M to allow incorporation of data from multiple experiments into the statistical analyses and figures, as raw optical density (OD) values were not always identical in different experiments.
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Loss-of-binding of mutant proteins will be associated with alterations in the level of expression of many genes in the cardiac network through haploinsufficiency, which will be a major contributor to the structural and functional heart defects found in human CHD patients.
To address the biochemical effects of missense variants on FABP7 and FABP5, we determined the binding potential of mutant proteins to ligands.
Drug binding affinities of mutant proteins were determined, and structures of QacR(E58Q) bound to Be, Dq, and MG and QacR(E57Q) bound to Dq and MG were determined.
We generated nine full-length proteins mutated at ionic and non-ionic polar residues between positions 83 and 96 corresponding to the F-helix and FG loop, and examined the heme binding properties, autoxidation rates, and catalytic activities of mutant proteins.
DOI: http://dx.doi.org/10.7554/eLife.03579.021 Residues involved in substrate binding were replaced and the transport activity of mutant proteins was measured in proteoliposomes.
Experiments of mutant proteins demonstrated that binding specificity is determined by the core basic region and the hinge region independently, and the two regions have an additive effect on DNA-binding specificity.
IgE binding of the mutant proteins was evaluated by in vitro studies using patients' sera.
We then studied the catalytic activities and DNA binding of these mutant proteins as well as some important functional protein interactions.
Finally, we show that one consequence shared by these mutations is the reduced binding of mutant Spt16 proteins across SRG1 and other highly transcribed genes.
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