The other experimental details have been described elsewhere.69 Appropriate blank corrections corresponding to curves obtained for samples containing only buffers were made in all spectra. Following the standard protocols used in our laboratories, the samples were prepared the day before and left over...
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In that complex, the single Trp residue of melittin preferentially binds to CaMC in full-length CaM, and CaMN interacts elsewhere; however, CaMN as an isolated fragment will bind to the Trp residue that is available because CaMC is absent. In the titrations of the FGF12ACaMBD biosensor ...
Proteins were digested with sequencing grade-modified trypsin (Roche Diagnostics) as described elsewhere.19 Tryptic peptides were extracted from gel pieces in 50% acetonitrile with 5% formic acid and dried using a vacuum centrifuge. Dried peptides were resuspended in 5% acetonitrile containing 0.1% ...
The DNA binding ability of all mutants was confirmed by titration experiments as described elsewhere10,11,12. For NMR analysis, the p53 gene corresponding to residues 313–393 of human p53 in pGEX-6P-1 was generated using a KOD-Plus-Mutagenesis Kit (TOYOBO, Osaka, Japan). 15N-labeled p53 ...
composed predominately of water, up to 70–80%, with the matrix forming component of the extracellular matrix (ECM) including a range of often highly post translationally modified proteins including collagens, proteoglycans such as aggrecan, laminins and fibronectin, reviewed in detail elsewhere15,...
0.25 % sodium deoxycholate, 150 mM NaCl, 1 mM EDTA, 1 mM PMSF, and 1 mM NaF, 1 mM Na3VO4,and protease inhibitor cocktail (Roche), and then resolved in SDS-polyacrylamide electrophoresis for immunoblotting. Quantitative PCR assays were carried out using SYBR Green as described elsewhere [...
The shorter 3′-UTRwas chosen for these studies because the first polyadenylation site ispredominantly used in differentiating C2C12 cells(23). To generate the AChE3′-UTR fragment without the AU-rich element (ARE), we employed aPCR-based protocol described elsewhere and used to delete the ARE ...
The other experimental parameters used in the experiments have been described elsewhere [48]. Appropriate blank corrections were made in all spectra. 2.4.2. Binding Experiments with PADI4 For the titration between either Impα3 or ΔImpα3 and intact PADI4, increasing amounts of the ...
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