Anti-microbial properties of histone H2A from skin secretions of rainbow trout, Oncorhynchus mykiss.

Skin exudates of rainbow trout contain a potent 13.6 kDa anti-microbial protein which, from partial internal amino acid sequencing, peptide mass fingerprinting, matrix-associated laser desorption/ionization MS and amino acid analysis, seems to be histone H2A, acetylated at the N-terminus. The protein, purified to homogeneity by ion-exchange and reversed-phase chromatography, exhibits powerful anti-bacterial activity against Gram-positive bacteria, with minimal inhibitory concentrations in the submicromolar range. Kinetic analysis revealed that at a concentration of 0.3 microM all test bacteria lose viability after 30 min incubation. Weaker activity is also displayed against the yeast Saccharomyces cerevisiae. The protein is salt-sensitive and has no haemolytic activity towards trout erythrocytes at concentrations below 0.3 microM. Reconstitution of the protein in a planar lipid bilayer strongly disturbs the membrane but does not form stable ion channels, indicating that its anti-bacterial activity is probably not due to pore-forming properties. This is the first report to show that, in addition to its classical function in the cell, histone H2A has extremely strong anti-microbial properties and could therefore help contribute to protection against bacterial invasion.

[1]  David Gottlieb,et al.  Mechanism of Action , 2012, Antibiotics.

[2]  S. C. Kim,et al.  Cathepsin D produces antimicrobial peptide parasin I from histone H2A in the skin mucosa of fish , 2002, FASEB journal : official publication of the Federation of American Societies for Experimental Biology.

[3]  M. Zasloff.  Antimicrobial peptides of multicellular organisms , 2002, Nature.

[4]  K. Buchmann.  Lectins in fish skin: do they play a role in host–monogenean interactions? , 2001, Journal of Helminthology.

[5]  C. Hawryshyn,et al.  Functional mapping of ultraviolet photosensitivity during metamorphic transitions in a salmonid fish, Oncorhynchus mykiss. , 2001, The Journal of experimental biology.

[6]  R C Richards,et al.  Histone H1: an antimicrobial protein of Atlantic salmon (Salmo salar). , 2001, Biochemical and biophysical research communications.

[7]  R. Hancock,et al.  Pleurocidin Coho Salmon with Lysozyme and Flounder Synergy of Histone-derived Peptides Of , 2001 .

[8]  S. Lovas,et al.  The antibacterial peptide pyrrhocoricin inhibits the ATPase actions of DnaK and prevents chaperone-assisted protein folding. , 2001, Biochemistry.

[9]  N. Orange,et al.  Isolation and characterization of novel glycoproteins from fish epidermal mucus: correlation between their pore-forming properties and their antibacterial activities. , 2000, Biochimica et biophysica acta.

[10]  Huey W. Huang,et al.  Action of antimicrobial peptides: two-state model. , 2000, Biochemistry.

[11]  S. Jones,et al.  Antibacterial proteins in rainbow trout, Oncorhynchus mykiss. , 2000, Fish & shellfish immunology.

[12]  Y. Shai,et al.  Mechanism of the binding, insertion and destabilization of phospholipid bilayer membranes by alpha-helical antimicrobial and cell non-selective membrane-lytic peptides. , 1999, Biochimica et biophysica acta.

[13]  D. N. Perkins,et al.  Probability‐based protein identification by searching sequence databases using mass spectrometry data , 1999, Electrophoresis.

[14]  J. Relf,et al.  Purification and characterization of a cysteine-rich 11.5-kDa antibacterial protein from the granular haemocytes of the shore crab, Carcinus maenas. , 1999, European journal of biochemistry.

[15]  R. Hancock,et al.  Salt-Resistant Alpha-Helical Cationic Antimicrobial Peptides , 1999, Antimicrobial Agents and Chemotherapy.

[16]  N. Orange,et al.  Pore-forming properties and antibacterial activity of proteins extracted from epidermal mucus of fish. , 1999, Comparative biochemistry and physiology. Part A, Molecular & integrative physiology.

[17]  C. B. Park,et al.  Parasin I, an antimicrobial peptide derived from histone H2A in the catfish, Parasilurus asotus , 1998, FEBS letters.

[18]  D. Barra,et al.  Experimental Infections of Rana esculenta with Aeromonas hydrophila: A Molecular Mechanism for the Control of the Normal Flora , 1998, Scandinavian journal of immunology.

[19]  R. Holle,et al.  Antimicrobial peptides melittin and cecropin inhibit replication of human immunodeficiency virus 1 by suppressing viral gene expression. , 1998, The Journal of general virology.

[20]  T. Arroll,et al.  Antimicrobial activity in the skin of the channel catfish Ictalurus punctatus: characterization of broad-spectrum histone-like antimicrobial proteins , 1998, Cellular and Molecular Life Sciences CMLS.

[21]  C. B. Park,et al.  Mechanism of action of the antimicrobial peptide buforin II: buforin II kills microorganisms by penetrating the cell membrane and inhibiting cellular functions. , 1998, Biochemical and biophysical research communications.

[22]  Brian Salmons,et al.  Expression of antimicrobial peptides has an antitumour effect in human cells. , 1998, Biochemical and biophysical research communications.

[23]  I. Hirono,et al.  Characterization and expression of c-type lysozyme cDNA from Japanese flounder (Paralichthys olivaceus). , 1997, Molecular marine biology and biotechnology.

[24]  Y. Kirino,et al.  Membrane permeabilization mechanisms of a cyclic antimicrobial peptide, tachyplesin I, and its linear analog. , 1997, Biochemistry.

[25]  G. Diamond,et al.  Isolation and Characterization of Pleurocidin, an Antimicrobial Peptide in the Skin Secretions of Winter Flounder* , 1997, The Journal of Biological Chemistry.

[26]  S J Ludtke,et al.  Membrane pores induced by magainin. , 1996, Biochemistry.

[27]  H. Jörnvall,et al.  Biochemical and antibacterial analysis of human wound and blister fluid. , 1996, European journal of biochemistry.

[28]  Y. Shai,et al.  A class of highly potent antibacterial peptides derived from pardaxin, a pore-forming peptide isolated from Moses sole fish Pardachirus marmoratus. , 1996, European journal of biochemistry.

[29]  C. B. Park,et al.  A novel antimicrobial peptide from Bufo bufo gargarizans. , 1996, Biochemical and biophysical research communications.

[30]  J. Thompson,et al.  CLUSTAL W: improving the sensitivity of progressive multiple sequence alignment through sequence weighting, position-specific gap penalties and weight matrix choice. , 1994, Nucleic acids research.

[31]  C. Murphy,et al.  Defensins are mitogenic for epithelial cells and fibroblasts , 1993, Journal of cellular physiology.

[32]  L. Clem,et al.  Temperature and teleost immune functions , 1992 .

[33]  K. V. van Holde,et al.  Nucleosome positioning is determined by the (H3-H4)2 tetramer. , 1991, Proceedings of the National Academy of Sciences of the United States of America.

[34]  M. Kashima.  H1 Histones Contribute to Candidacidal Activities of Human Epidermal Extract , 1991, The Journal of dermatology.

[35]  R. Jackson,et al.  Ultrasensitive assays for endogenous antimicrobial polypeptides. , 1991, Journal of immunological methods.

[36]  E. Myers,et al.  Basic local alignment search tool. , 1990, Journal of molecular biology.

[37]  H. Schägger,et al.  Tricine-sodium dodecyl sulfate-polyacrylamide gel electrophoresis for the separation of proteins in the range from 1 to 100 kDa. , 1987, Analytical biochemistry.

[38]  N. Heintz,et al.  The primary structure and expression of four cloned human histone genes. , 1983, Nucleic acids research.

[39]  J. Raa,et al.  Skin mucus protease from rainbow trout, Salmo gairdneri Richardson, and its biological significance , 1983 .

[40]  M. M. Bradford.  A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. , 1976, Analytical biochemistry.

[41]  M Montal,et al.  Formation of bimolecular membranes from lipid monolayers and a study of their electrical properties. , 1972, Proceedings of the National Academy of Sciences of the United States of America.

[42]  J. G. Hirsch.  BACTERICIDAL ACTION OF HISTONE , 1958, The Journal of experimental medicine.

[43]  T. Yano.  3 – The Nonspecific Immune System: Humoral Defense , 1996 .

[44]  C. Bevins.  Antimicrobial peptides as agents of mucosal immunity. , 1994, Ciba Foundation symposium.