Peptide-based analysis of amino acid sequences important to the biological activity of eosinophil granule major basic protein

Larry L. Thomas, Hirokazu Kubo, David J. Loegering, Kristine Spillard, Arthur J. Weaver, Daniel J. McCormick, Catherine Weiler, Gerald J. Gleich

Research output: Contribution to journalArticle

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Abstract

Synthetic peptides corresponding to amino acid sequences in eosinophil granule major basic protein (MBP) were evaluated for cytotoxic activity toward K562 cells and for ability to stimulate basophil mediator release. Results obtained using 14 peptides spanning the 117-amino acid sequence of MBP in overlapping fashion indicated that the activities mapped to peptide sequences near the amino and carboxy termini of MBP. The activity of these regions was confirmed using two peptides corresponding to MBP residues 18-45 and 89-117. A 20-h incubation with 5 μM peptide 18-45 or peptide 89-117 caused approximately the same levels ( > 60%) of cytotoxicity in K562 cells as 5 μM MBP. Similarly, a 30-min incubation with peptides 18-44 and 89-117 stimulated basophil histamine release in a concentration-dependent manner over the range of 5-20 μM. The level of release stimulated by 20 μM peptide 89-117 approached that stimulated by 2 μM MBP. A 20 μM concentration of peptide 89-117 also stimulated leukotriene C4 (LTC4) production by the basophils. Neither peptide 18-45 nor peptide 89-117 was cytotoxic for basophils under the experimental conditions for histamine and LTC4 release, as determined by 51Cr release. These results indicate that two MBP peptide sequences, including one (89-117) that contains a unique carbohydrate-binding region, share the biologic activities of MBP.

Original languageEnglish (US)
Pages (from-to)175-181
Number of pages7
JournalImmunology Letters
Volume78
Issue number3
DOIs
StatePublished - Oct 1 2001

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Keywords

  • Basophil
  • Cell activation
  • Cytotoxicity
  • Eosinophil
  • Major basic protein

ASJC Scopus subject areas

  • Immunology and Allergy
  • Immunology

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