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We identify a mutant predicted to lead to increased interfacial hydrophobic contacts (N92F) that shows markedly increased self-association properties on native gradient gels.
Specifically, the T14M/T15K mutant, predicted to impair ATP hydrolysis, as well as the W125D substitution, which deforms and adds charge to the catalytic cleft, exhibited virtually complete abrogation of kinase function.
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Generation of triple lysine mutants predicted to completely disrupt the putative TMD allowed us to examine TMD function in the full-length proteins.
In our human DC experiments, mutants predicted to result in a non-mannose capped LAM (Rv2181, Rv1635c, Rv1565c) all had a fitness cost both 3 days and 7 days post-infection (Additional file 4).
Using genetic rescue approaches, CPX mutants predicted to prevent the formation of the zig-zag array (CPX GPGP) disrupt the ability of CPX to clamp spontaneous release, while mutations predicted to enhance the CPXacc t-SNARE interaCPXacc t-SNARElamp mutatinteractiont a strong clamping ability (Cho et al., 2014).
Measured by flow cytometry, the mutants predicted to enforce pairing within each monomer functioned better than those enforcing the interaction between the monomers; although none of the four mutants silenced EGFP as efficiently as the canonical repeat A 2-mer, suggesting that there may be more complex structures involved.
The M4 mutant is predicted to lose the shared GATA1/GATA2 site and the downstream SP1 site, The M6 mutant loses a predicted binding site for zeste homolog/TCERG1.
Synthetic peptides corresponding to mutant proteins predicted to arise from MSI-associated frameshift mutations have been shown to be immunogenic.
Therefore cone photoreceptor degeneration caused by GCAP1-Y99C mutare are predicted to also respond favorably to prolonged light exposure.
All other codons of the gene remain wild-type, therefore the mutant is predicted to be non-polar.
The secondary structure of the 152 167 peptide and S158E mutant is predicted to contain a beta-strand (DNAMan software, Lynnon Biosoft).
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