Identification of human cysteine-rich secretory protein 3 (CRISP-3) as a matrix protein in a subset of peroxidase-negative granules of neutrophils and in the granules of eosinophils

J Leukoc Biol. 2002 Sep;72(3):462-9.


Cysteine-rich secretory protein 3 (CRISP-3; also known as SGP28) was originally discovered in human neutrophilic granulocytes. We have recently developed a sensitive sandwich enzyme-linked immunosorbent assay for CRISP-3 and demonstrated the presence of CRISP-3 in exocrine secretions. To investigate the subcellular localization and mobilization of CRISP-3 in human neutrophils, we performed subcellular fractionation of resting and activated neutrophils on three-layer Percoll density gradients, release-studies of granule proteins in response to different secretagogues, and double-labeling immunogold electron microscopy. CRISP-3 was found to be localized in a subset of granules with overlapping characteristics of specific and gelatinase granules and mobilized accordingly, thus confirming the hypothesis that peroxidase-negative granules exist as a continuum from specific to gelatinase granules regarding protein content and mobilization. CRISP-3 was found to be a matrix protein, which is stored in granules as glycosylated and as unglycosylated protein. The subcellular distribution of the two forms of CRISP-3 was identical. In addition, CRISP-3 was found as a granule protein in eosinophilic granulocytes. The presence of CRISP-3 in peroxidase-negative granules of neutrophils, in granules of eosinophils, and in exocrine secretions indicates a role in the innate host defense.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Cell Fractionation
  • Centrifugation, Density Gradient
  • Cytoplasmic Granules / chemistry*
  • Cytoplasmic Granules / metabolism
  • Cytoplasmic Granules / ultrastructure
  • Eosinophil Peroxidase
  • Eosinophils / chemistry*
  • Eosinophils / drug effects
  • Eosinophils / ultrastructure
  • Gelatinases / metabolism
  • Glycosylation
  • Humans
  • Ionomycin / pharmacology
  • Lactoferrin / metabolism
  • Microscopy, Immunoelectron
  • N-Formylmethionine Leucyl-Phenylalanine / pharmacology
  • Peroxidase / metabolism
  • Peroxidases / analysis
  • Protein Processing, Post-Translational
  • Salivary Proteins and Peptides / analysis
  • Salivary Proteins and Peptides / chemistry
  • Salivary Proteins and Peptides / physiology*
  • Seminal Plasma Proteins / analysis
  • Seminal Plasma Proteins / chemistry
  • Seminal Plasma Proteins / physiology*
  • Subcellular Fractions / chemistry
  • Tetradecanoylphorbol Acetate / pharmacology
  • Zymosan / pharmacology


  • CRISP3 protein, human
  • Salivary Proteins and Peptides
  • Seminal Plasma Proteins
  • Ionomycin
  • N-Formylmethionine Leucyl-Phenylalanine
  • Zymosan
  • Eosinophil Peroxidase
  • Peroxidases
  • Peroxidase
  • Lactoferrin
  • Gelatinases
  • Tetradecanoylphorbol Acetate