title: Novel haemoglobin-derived antimicrobial peptides from chicken (Gallus gallus) blood: purification, structural aspects and biological activity.
abstract: OBJECTIVE
  To purify and characterize antimicrobial peptides derived from the acid extract of Gallus gallus blood cells.
  
  
  RESULTS
  Two polypeptides (i.e. CHb-1 and CHb-2) with antibacterial activity were detected in the acidic extract of blood cells from chicken (G. gallus). The isolated peptides that possessed a potent antibacterial activity were purified using a two-step chromatography procedure that involved solid-phase extraction of a total protein/peptide extract followed by thin fractionation by reversed-phase high performance liquid chromatography (RP-HPLC). The molecular masses of the purified peptides were similar and were 4824·4 and 4825·2 Da, which have been measured by matrix-assisted laser desorption/ionization mass spectrometry (MALDI TOF MS). Their amino acid sequences were determined by Edman degradation and showed that the peptides were fully identical to the two fragments of G. gallus α-haemoglobin localized into different subunits (A and D respectively). The peptides were active in micromolar concentrations against Gram-negative Escherichia coli K12 TG1. Using the 1-N-phenylnaphthylamine, the FITC-dextran labelled probes and the live/dead staining allowed to show the hemocidin mode of action and estimate the pore size.
  
  
  CONCLUSIONS
  In this study, for the first time, α-haemoglobin from chicken (G. gallus) has been investigated as a donor of the two high homologous native peptide fragments that possess potent antibacterial activity in vitro. These are membrane-active peptides and their mechanism of action against E. coli involves a toroidal pore formation.
  
  
  CONCLUSIONS
  The obtained results expand the perception of the role of haemoglobin in a living system, describing it as a source of multifunction substances. Additionally, the data presented in this paper may contribute to the development of new, cost-effective, antimicrobial agents.
authors: Institute of Cellular and Intracellular Symbiosis, RAS, Orenburg, Russia.
authors: A S
authors: AS
authors: Vasilchenko
authors: Shemyakin and Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of Sciences, Moscow, Russia.|Gause Institute of New Antibiotics, Moscow, Russia.
authors: E A
authors: EA
authors: Rogozhin
authors: Orenburg State University, Orenburg, Russia.
authors: A V
authors: AV
authors: Vasilchenko
authors: Institute of Cellular and Intracellular Symbiosis, RAS, Orenburg, Russia.
authors: O L
authors: OL
authors: Kartashova
authors: Institute of Cellular and Intracellular Symbiosis, RAS, Orenburg, Russia.|Orenburg State Agrarian University, Orenburg, Russia.
authors: M V
authors: MV
authors: Sycheva
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D000900:Anti-Bacterial Agents / Q000737:chemistry / Q000302:isolation & purification / Q000494:pharmacology*; D002645:Chickens; D004926:Escherichia coli / Q000187:drug effects; D006454:Hemoglobins / Q000737:chemistry / Q000302:isolation & purification / Q000494:pharmacology*; D008970:Molecular Weight; D010446:Peptide Fragments / Q000737:chemistry / Q000302:isolation & purification / Q000494:pharmacology*
chemical_list: D000900:Anti-Bacterial Agents; C000615786:CHb-1 peptide, chicken; C000615787:CHb-2 peptide, chicken; D006454:Hemoglobins; D010446:Peptide Fragments; C070077:alpha(A) globin
keywords: 
  Escherichia coli
  ; 
  Gallus gallus
  ; antimicrobial peptides; hemocidin; mode of action