Primary sequence, together with other factors, influence peptide deimination by peptidylarginine deiminase-4
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Maria E. Stensland
Abstract
Enzymes of the peptidylarginine deiminase (PAD) family catalyze the posttranslational deimination of polypeptide-bound arginine residues. Here, we report the selection of peptide substrates by PAD-4, an isoform thought to be involved in the pathogenesis of rheumatoid arthritis. First, we investigated whether PAD-4-mediated deimination is influenced by the nature of amino acid residues flanking the targeted arginine. Using two peptide substrates, residues in positions -2, -1, +1, and +2 relative to the central arginine targeted by PAD-4 were systematically replaced by all natural l-amino acids except cysteine. Each peptide was treated with recombinant human PAD-4 and deimination was analyzed by matrix-assisted laser desorption-ionization time-of-flight mass spectrometry. In all four flanking positions, amino acids which positively or negatively influenced deimination were identified. We next designed peptides with expected high or low deimination rates and determined their Km and kcat values. These peptides showed PAD-4 substrate behavior as predicted, demonstrating that residues flanking the targeted arginine are important for deimination. Further truncation of peptide substrates suggested additional effects on deimination by residues outside the -2 to +2 region. Finally, we observed that a methylated lysine residue flanking the targeted arginine influences PAD-4-mediated deimination, also suggesting that posttranslational modifications can affect substrate efficiency.
©2009 by Walter de Gruyter Berlin New York
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Articles in the same Issue
- Minireview
- Functional genetic mouse models: promising tools for investigation of the proteolytic internet
- Protein Structure and Function
- Primary sequence, together with other factors, influence peptide deimination by peptidylarginine deiminase-4
- Mercury and cadmium trigger expression of the copper importer Ctr1B, which enables Drosophila to thrive on heavy metal-loaded food
- Cell Biology and Signaling
- Interferon-γ-mediated pathways and in vitro PBMC proliferation in HIV-infected patients
- Exploring the pathogenesis of renal cell carcinoma: pathway and bioinformatics analysis of dysregulated genes and proteins
- Aptamers selected against the unglycosylated EGFRvIII ectodomain and delivered intracellularly reduce membrane-bound EGFRvIII and induce apoptosis
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- Overexpression of Pygopus2 protects HeLa cells from vinblastine-induced apoptosis
- Proteolysis
- Selectivity of propeptide-enzyme interaction in cathepsin L-like cysteine proteases
- Murine and human cathepsin B exhibit similar properties: possible implications for drug discovery
- Novel Techniques
- Isotope tracing enhancement of chemiluminescence assays for nitric oxide research
