Vertical-scanning mutagenesis of amino acids in a model N-myristoylation motif reveals the major amino-terminal sequence requirements for protein N-myristoylation.

@article{Utsumi2004VerticalscanningMO,
  title={Vertical-scanning mutagenesis of amino acids in a model N-myristoylation motif reveals the major amino-terminal sequence requirements for protein N-myristoylation.},
  author={Toshihiko Utsumi and Kengo Nakano and Takeshi Funakoshi and Yoshiyuki Kayano and Sayaka Nakao and Nagisa Sakurai and H Iwata and Rumi Ishisaka},
  journal={European journal of biochemistry},
  year={2004},
  volume={271 4},
  pages={
          863-74
        }
}
In order to determine the amino-terminal sequence requirements for protein N-myristoylation, site-directed mutagenesis of the N-terminal region was performed using tumor necrosis factor (TNF) mutants as model substrate proteins. Subsequently, the susceptibility of these mutants to protein N-myristoylation was evaluated by metabolic labeling in an in vitro translation system using rabbit reticulocyte lysate. A TNF mutant having the sequence MGAAAAAAAA at its N-terminus was used as the starting… 
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TLDR
The amino acid residue penultimate to the N-terminal Gly residue strongly affected two cotranslational protein modifications, N-myristoylation andN-acetylation, and the amino acid requirements at this position for these two modifications were significantly affected by downstream residues.
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TLDR
Indications that, at least within a complete substrate protein, the N-terminal 17 protein residues experience different types of variability restrictions are found, which are proposed for the design of NMT inhibitors in pathogenic fungal and protozoan systems.
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TLDR
A scan of public protein databases revealed new potential NMT targets for which the myristoyl modification may be of critical importance for biological function and a version of the predictor is proposed that identifies a number of proteolytic protein processing sites at internal glycine residues and that evaluates possible N-terminalMyristoylation of the protein fragments.
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TLDR
A survey of 26 derivatives of this inhibitor confirmed the important stereochemical requirements for the N-terminal amine, the β-hydroxyl of Ser5, and the ε-amino group of Lys6, and established that a simple alkyl backbone can maintain an appropriate distance between three elements critical for recognition by the fungal enzyme’s peptide-binding site.
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  • Biology, Chemistry
    Proceedings of the National Academy of Sciences of the United States of America
  • 1987
TLDR
It is established that structural information important for NMT-ligand interaction exists beyond the first two amino acids in peptide substrates and that the side chains of residue 5 play a critical role in the binding of substrates to this enzyme.
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TLDR
Heterologous fusion proteins containing the first 10 amino acids of Gi1 alpha and Gs alpha using tumor necrosis factor as a model protein determined their ability to incorporate palmitate using in vitro and in vivo expression systems and indicated the Met-Gly-Cys motif found in G-protein alpha subunits itself is not sufficient to direct palmitoylation even if Gly-2 is myristoylated after removal of initiating Met.
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TLDR
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TLDR
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TLDR
A heretofore unappreciated level of genetic complexity underlying the enzymology of N-terminal myristoylation is demonstrated and it is suggested that the specific inhibition or regulation of either NMT in vivo may in turn allow for the selective control of particular myristylation-dependent cellular functions.
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