Using the mutagenic PCR method, random nucleotides in the binding site were substituted with different nucleotides to impair HOXD10 binding.
Strikingly, several amino acid residues known to be involved in TPP binding were substituted in the MmTKL proteins compared to the corresponding residues of ScTKL1.
Mutagenesis was performed using a QuikChange Site-Directed Mutagenesis Kit (Stratagene, La Jolla, CA, USA), and all the adenines of miRNA-binding sequences were substituted with thymines.
Comparative analysis revealed expanded numbers of bZIP candidates in oomycetes, especially the Phytophthora species, due to the expansion of several novel bZIP classes whose highly conserved asparagines in basic DNA-binding regions were substituted by other residues such as cysteine.
To confirm this, we used an sRNA-binding deficient Ago2 mutant (Y311A/F312A or YA/FA), in which two conserved residues important for sRNA-binding were substituted with alanine.
Three Trp residues in the ELC binding region of MYHHead3_As substituted for nonaromatic aa in scallop myosins, while 2 Trp residues in the RLC binding region in scallop myosins were substituted by two other aromatic aa in MYHHead3_As.
In order to clarify the impact of ubiquitination on the endocytosis of L1, I constructed a ubiquitination-deficient mutant of L1, L1K11R, in which 11 Lys residues, two of which constitute a putative ERM-binding motif (Fig. 1A), were substituted with Arg residues, because ubiquitination occurs predominantly at the ε-amino group of the lysine residue within a substrate.
Although none of the synthetic peptides used in this study were substituted at MHC binding positions, we explored whether the analogue glycopeptides were able to elicit an MHC restricted immune T-cell response.
Active components of the zeolite were determined by means of X-ray diffraction analysis, particle form (plate-shaped) and size (11.82 μm) were determined by means of SEM. 5 batches of specimens were made with zeolite content ranging from 0 to 10% (the binding material was substituted with natural zeolite).
The absence of S. cerevisiae binding sites in the distantly related species could be the result of a more complex model whereby one binding site is substituted for another site bound by a different transcription factor.
