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PMID:2500436

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Citation

Wilde, CG, Griffith, JE, Marra, MN, Snable, JL and Scott, RW (1989) Purification and characterization of human neutrophil peptide 4, a novel member of the defensin family. J. Biol. Chem. 264:11200-3

Abstract

Primary (azurophil) granules of neutrophils contain proteins which play a major role in the killing and digestion of bacteria in the phagolysosome. We have isolated and characterized a novel antimicrobial peptide from the azurophil granule fraction of discontinuous Percoll gradients. We have named this peptide human neutrophil peptide 4 (HNP-4) based on its structural similarity to a group of antimicrobial polypeptides known as defensins (HNP 1-3). Using size exclusion and reverse-phase high performance liquid chromatography, HNP-4 was purified to homogeneity as judged by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and amino-terminal sequence analysis. The amino acid sequence determined from isolated HNP-4 and from tryptic fragments of reduced and alkylated peptide is: NH2-Val-Cys-Ser-Cys-Arg-Leu-Val-Phe-Cys-Arg-Arg-Thr-Glu- Leu-Arg-Val-Gly-Asn-Cys-Leu-Ile-Gly-Gly-Val-Ser-Phe-Thr-Tyr-Cys-Cys-Thr- Arg-Val - COOH. Based on this sequence, HNP-4 has a calculated molecular weight of 3715 and a theoretical pI of 8.61. HNP-4 shows structural similarity to the family of three human defensins. HNP-4 and the defensins have identical cysteine backbones and, like the defensins, HNP-4 is rich in arginine (15.2 mol %). However, the amino acids at 22 of the 33 positions differ between HNP-4 and human defensins. Further, HNP-4 is significantly more hydrophobic than the defensins, as determined by its retention time on reverse-phase high performance liquid chromatography. In vitro, purified HNP-4 was shown to kill Escherichia coli, Streptococcus faecalis, and Candida albicans. Compared to a mixture of the other human defensins, HNP-4 was found to be approximately 100 times more potent against E. coli and four times more potent against both S. faecalis and C. albicans.

Links

PubMed

Keywords

Amino Acid Sequence; Amino Acids/analysis; Blood Bactericidal Activity; Blood Proteins/isolation & purification; Blood Proteins/physiology; Candida albicans; Chromatography, Gel; Chromatography, High Pressure Liquid; Cytoplasmic Granules/analysis; Electrophoresis, Polyacrylamide Gel; Enterococcus faecalis; Escherichia coli; Humans; Isoelectric Point; Molecular Sequence Data; Molecular Weight; Neutrophils/analysis; Neutrophils/ultrastructure; alpha-Defensins

Significance

Annotations

Gene product Qualifier GO Term Evidence Code with/from Aspect Extension Notes Status

HUMAN:DEF4

located_in

GO:0042582: azurophil granule

ECO:0000314: direct assay evidence used in manual assertion

C

  • part_of:(CL:0000775)

Seeded From UniProt

complete

HUMAN:DEF4

GO:0044194: cytolytic granule

ECO:0000314:

C

microbicidal assays, figure 1 in experimental procedures. purified and isolated granules and showed that the proteins came from the granules.

complete

HUMAN:DEF4

involved_in

GO:0061844: antimicrobial humoral immune response mediated by antimicrobial peptide

ECO:0000314: direct assay evidence used in manual assertion

P

Seeded From UniProt

complete

HUMAN:DEF4

involved_in

GO:0050832: defense response to fungus

ECO:0000314: direct assay evidence used in manual assertion

P

Seeded From UniProt

complete

HUMAN:DEF4

GO:0042742: defense response to bacterium

ECO:0000314:

P

table 2

complete

HUMAN:DEF4

involved_in

GO:0019732: antifungal humoral response

ECO:0000314: direct assay evidence used in manual assertion

P

Seeded From UniProt

complete

HUMAN:DEF4

involved_in

GO:0019730: antimicrobial humoral response

ECO:0000314: direct assay evidence used in manual assertion

P

Seeded From UniProt

complete

HUMAN:DEF4

involved_in

GO:0019731: antibacterial humoral response

ECO:0000314: direct assay evidence used in manual assertion

P

Seeded From UniProt

complete

HUMAN:DEF4

involved_in

GO:0045087: innate immune response

ECO:0000314: direct assay evidence used in manual assertion

P

Seeded From UniProt

complete

HUMAN:DEF4

involved_in

GO:0050829: defense response to Gram-negative bacterium

ECO:0000314: direct assay evidence used in manual assertion

P

Seeded From UniProt

complete

HUMAN:DEF4

involved_in

GO:0050830: defense response to Gram-positive bacterium

ECO:0000314: direct assay evidence used in manual assertion

P

Seeded From UniProt

complete

HUMAN:DEF4

part_of

GO:0042582: azurophil granule

ECO:0000314: direct assay evidence used in manual assertion

C

part_of:(CL:0000775)

Seeded From UniProt

complete


See also

References

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