Thrombocidins, microbicidal proteins from human blood platelets, are C-terminal deletion products of CXC chemokines.
Antibacterial proteins are components of the innate immune system found in many organisms and produced by a variety of cell types. Human blood platelets contain a number of antibacterial proteins in their alpha-granules that are released upon thrombin activation. The present study was designed to purify these proteins obtained from human platelets and to characterize them chemically and biologically. Two antibacterial proteins were purified from platelet granules in a two-step protocol using cation exchange chromatography and continuous acid urea polyacrylamide gel electrophoresis and were designated thrombocidin (TC)-1 and TC-2. Characterization of these proteins using mass spectrometry and N-terminal sequencing revealed that TC-1 and TC-2 are variants of the CXC chemokines neutrophil-activating peptide-2 and connective tissue-activating peptide-III, respectively. TC-1 and TC-2 differ from these chemokines by a C-terminal truncation of 2 amino acids. Both TCs, but not neutrophil-activating peptide-2 and connective tissue-activating peptide-III, were bactericidal for Bacillus subtilis, Escherichia coli, Staphylococcus aureus, and Lactococcus lactis and fungicidal for Cryptococcus neoformans. Killing of B. subtilis by either TC appeared to be very rapid. Because TCs were unable to dissipate the membrane potential of L. lactis, the mechanism of TC-mediated killing most probably does not involve pore formation.
Krijgsveld
J
J
Department of Medical Microbiology, Academic Medical Center, University of Amsterdam, 1105 AZ Amsterdam, The Netherlands.
Zaat
S A
SA
Meeldijk
J
J
van Veelen
P A
PA
Fang
G
G
Poolman
B
B
Brandt
E
E
Ehlert
J E
JE
Kuijpers
A J
AJ
Engbers
G H
GH
Feijen
J
J
Dankert
J
J
D000595:Amino Acid Sequence; D000900:Anti-Bacterial Agents / Q000737:chemistry* / Q000494:pharmacology; D000935:Antifungal Agents / Q000737:chemistry; D001419:Bacteria / Q000187:drug effects; D001792:Blood Platelets / Q000737:chemistry*; D001798:Blood Proteins / Q000737:chemistry*; D018925:Chemokines; D019743:Chemokines, CXC / Q000737:chemistry*; D006801:Humans; D007700:Kinetics; D013058:Mass Spectrometry; D008564:Membrane Potentials / Q000187:drug effects; D008969:Molecular Sequence Data; D009504:Neutrophils / Q000737:chemistry; D020539:Sequence Analysis, Protein; D001620:beta-Thromboglobulin
D000900:Anti-Bacterial Agents; D000935:Antifungal Agents; D001798:Blood Proteins; D018925:Chemokines; D019743:Chemokines, CXC; C061615:PPBP protein, human; D001620:beta-Thromboglobulin