Modified peptide nucleic acids are internalized in mouse macrophages RAW 264.7 and inhibit inducible nitric oxide synthase

FEBS Letters

Millo, Enrico¹  Benatti, Umberto¹  Giovine, Marco¹  Pozzolini, Marina¹  Gasparini, Anna¹  Damonte, Gianluca¹  Scarfi, Sonia¹ 

[1] Department of Experimental Medicine, Biochemistry Section, University of Genoa, Viale Benedetto XV, 1 16132 Genoa, Italy

关键词: Peptide nucleic acid;    Inducible nitric oxide synthase;    RAW 267.4;    Internalization;    Boc;    tert-butoxycarbonyl;    ESMS;    electrospray mass spectrometry;    iNOS;    inducible nitric oxice synthase;    LPS;    lipopolysaccharide;    MBHA-PS;    4-methyl-benzhydril-amine-polystyrene;    PNA;    peptide nucleic acid;    RP-HPLC;    reverse phase high performance liquid chromatography;    TFA;    trifluoroacetic acid;   

DOI  :  10.1016/S0014-5793(99)00589-X

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

Overexpression of inducible nitric oxide synthase causes the production of high levels of nitric oxide, which, under pathological conditions, leads to immunosuppression and tissue damage. The results recently obtained using peptide nucleic acids, rather than traditional oligonucleotides as antigen and antisense molecules, prompted us to test their efficacy in the regulation of nitric oxide production, thereby overcoming the obstacle of cellular internalization. The cellular permeability of four inducible nitric oxide synthase antisense peptide nucleic acids of different lengths was evaluated. These peptide nucleic acids were covalently linked to a hydrophobic peptide moiety to increase internalization and to a tyrosine to allow selective ¹²⁵I radiolabelling. Internalization experiments showed a 3–25-fold increase in the membrane permeability of the modified peptide nucleic acids with respect to controls. inducible nitric oxide synthase inhibition experiments on intact stimulated macrophages RAW 264.7 after passive permeation of the two antisense peptide nucleic acids 3 and 4 demonstrated a significant decrease (43–44%) in protein enzymatic activity with respect to the controls. These data offer a basis for developing a good alternative to conventional drugs directed against inducible nitric oxide synthase overexpression.

【 授权许可】

Unknown