Antibacterial peptides in hemocytes and hematopoietic tissue from freshwater crayfish Pacifastacus leniusculus: characterization and expression pattern.
A 14 amino acid residues proline/arginine-rich antibacterial peptide designated as astacidin 2 was purified and characterized from hemocytes of the freshwater crayfish, Pacifastacus leniusculus. Astacidin 2 has a broad range of antibacterial activity against both Gram-positive and Gram-negative bacteria. The primary sequence of astacidin 2 is RPRPNYRPRPIYRP with an amidated C-terminal and the molecular mass is 1838Da determined by mass spectrometry. Furthermore, the cDNA of three different crustin antibacterial homologs were isolated from a crayfish hemocyte EST library. RT-PCR was used to analyze the expression of the genes coding for astacidin 2 and P. leniusculus crustins (Plcrustin) 1-3 after bacterial challenge. The expression of Plcrustin1 was upregulated in both hemocytes and hematopoietic tissue after challenge with Gram-negative Escherichia coli or Acinetobacter ssp. non pathogenic bacteria as well as by a Gram negative crayfish pathogenic bacterium (Aeromonas hydrophila). The PlCrustin3 transcript was only upregulated after inoculation with the non-pathogenic Acinetobacter ssp. while there was no change in expression of Plcrustin2 or astacidin 2 following a bacterial challenge.
Jiravanichpaisal
Pikul
P
Department of Comparative Physiology, Uppsala University, Norbyvägen 18A, SE-752 36, Sweden.
Lee
So Young
SY
Kim
Young-A
YA
Andrén
Tove
T
Söderhäll
Irene
I
D000595:Amino Acid Sequence; D000818:Animals; D023181:Antimicrobial Cationic Peptides / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism / Q000494:pharmacology; D003400:Astacoidea / Q000235:genetics* / Q000276:immunology / Q000378:metabolism; D001410:Bacillus megaterium / Q000187:drug effects; D001483:Base Sequence; D002852:Chromatography, Ion Exchange; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000737:chemistry / Q000235:genetics; D004926:Escherichia coli / Q000187:drug effects; D020869:Gene Expression Profiling; D006090:Gram-Negative Bacteria / Q000187:drug effects; D006094:Gram-Positive Bacteria / Q000187:drug effects; D006413:Hematopoietic System / Q000378:metabolism*; D006434:Hemocytes / Q000187:drug effects / Q000378:metabolism*; D006458:Hemolymph / Q000187:drug effects / Q000378:metabolism*; D043143:Hepatopancreas / Q000378:metabolism; D008070:Lipopolysaccharides / Q000494:pharmacology; D008826:Microbial Sensitivity Tests; D008969:Molecular Sequence Data; D021382:Protein Sorting Signals / Q000235:genetics; D017422:Sequence Analysis, DNA; D017386:Sequence Homology, Amino Acid; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
D023181:Antimicrobial Cationic Peptides; D018076:DNA, Complementary; D008070:Lipopolysaccharides; D021382:Protein Sorting Signals; C519609:RPRPNYRPRPIYRP peptide