title: Protein cross-linking by chlorinated polyamines and transglutamylation stabilizes neutrophil extracellular traps.
abstract: Neutrophil extracellular trap (NET) ejected from activated dying neutrophils is a highly ordered structure of DNA and selected proteins capable to eliminate pathogenic microorganisms. Biochemical determinants of the non-randomly formed stable NETs have not been revealed so far. Studying the formation of human NETs we have observed that polyamines were incorporated into the NET. Inhibition of myeloperoxidase, which is essential for NET formation and can generate reactive chlorinated polyamines through hypochlorous acid, decreased polyamine incorporation. Addition of exogenous primary amines that similarly to polyamines inhibit reactions catalyzed by the protein cross-linker transglutaminases (TGases) has similar effect. Proteomic analysis of the highly reproducible pattern of NET components revealed cross-linking of NET proteins through chlorinated polyamines and ɛ(γ-glutamyl)lysine as well as bis-γ-glutamyl polyamine bonds catalyzed by the TGases detected in neutrophils. Competitive inhibition of protein cross-linking by monoamines disturbed the cross-linking pattern of NET proteins, which resulted in the loss of the ordered structure of the NET and significantly reduced capacity to trap bacteria. Our findings provide explanation of how NETs are formed in a reproducible and ordered manner to efficiently neutralize microorganisms at the first defense line of the innate immune system.
authors: Department of Biochemistry and Molecular Biology, University of Debrecen, Debrecen, Hungary.
authors: Krisztián
authors: K
authors: Csomós
authors: Department of Biochemistry and Molecular Biology, University of Debrecen, Debrecen, Hungary.
authors: Endre
authors: E
authors: Kristóf
authors: Department of Biochemistry and Molecular Biology, University of Debrecen, Debrecen, Hungary.
authors: Bernadett
authors: B
authors: Jakob
authors: Department of Biophysics and Cell Biology, University of Debrecen, Debrecen, Hungary.
authors: István
authors: I
authors: Csomós
authors: Department of Computer Graphics and Image Processing, University of Debrecen, Debrecen, Hungary.
authors: György
authors: G
authors: Kovács
authors: Department of Biochemistry and Molecular Biology, University of Debrecen, Debrecen, Hungary.
authors: Omri
authors: O
authors: Rotem
authors: Department of Biochemistry and Molecular Biology, University of Debrecen, Debrecen, Hungary.
authors: Judit
authors: J
authors: Hodrea
authors: Division of Clinical Laboratory Science, University of Debrecen, Debrecen, Hungary.|Thrombosis and Hemostasis Research Group of the Hungarian Academy of Sciences, University of Debrecen, Debrecen, Hungary.
authors: Zsuzsa
authors: Z
authors: Bagoly
authors: Division of Clinical Laboratory Science, University of Debrecen, Debrecen, Hungary.|Thrombosis and Hemostasis Research Group of the Hungarian Academy of Sciences, University of Debrecen, Debrecen, Hungary.
authors: Laszlo
authors: L
authors: Muszbek
authors: Department of Biochemistry and Molecular Biology, University of Debrecen, Debrecen, Hungary.
authors: Zoltán
authors: Z
authors: Balajthy
authors: Department of Biochemistry and Molecular Biology, University of Debrecen, Debrecen, Hungary.
authors: Éva
authors: É
authors: Csősz
authors: Department of Biochemistry and Molecular Biology, University of Debrecen, Debrecen, Hungary.
authors: László
authors: L
authors: Fésüs
mesh_terms: D003432:Cross-Linking Reagents / Q000494:pharmacology*; D065206:Extracellular Traps / Q000187:drug effects / Q000378:metabolism*; D054879:Halogenation; D006801:Humans; D009504:Neutrophils / Q000187:drug effects / Q000378:metabolism*; D011073:Polyamines / Q000494:pharmacology*; D011506:Proteins / Q000378:metabolism*; D040901:Proteomics; D013211:Staphylococcus aureus / Q000187:drug effects / Q000378:metabolism; D011503:Transglutaminases / Q000378:metabolism*
chemical_list: D003432:Cross-Linking Reagents; D011073:Polyamines; D011506:Proteins; D011503:Transglutaminases