Identification of leukocyte cationic proteins that interact with ceruloplasmin

Proteins from leukocytes were investigated for their ability to interact with ceruloplasmin (Cp), a copper-containing glycoprotein of human plasma. Extract from leukocytes was subjected to affinity chromatography on Cp-Sepharose, after which proteins were eluted from the resin with 0.5 M NaCl in Tri...

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Bibliographic Details
Published in:Biochemistry (Moscow) 2007-08, Vol.72 (8), p.872-877
Main Authors: Sokolov, A V, Pulina, M O, Ageeva, K V, Runova, O L, Zakharova, E T, Vasil'ev, V B
Format: Article
Language:English
Subjects:
Cations - chemistry
Cations - isolation & purification
Cations - metabolism
Ceruloplasmin - chemistry
Ceruloplasmin - metabolism
Chromatography, Affinity
Humans
Immunoproteins - chemistry
Immunoproteins - isolation & purification
Immunoproteins - metabolism
Leukocytes - chemistry
Leukocytes - metabolism
Mass Spectrometry
Peptides
Protein Binding - physiology
Proteins
Sodium chloride
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Summary:Proteins from leukocytes were investigated for their ability to interact with ceruloplasmin (Cp), a copper-containing glycoprotein of human plasma. Extract from leukocytes was subjected to affinity chromatography on Cp-Sepharose, after which proteins were eluted from the resin with 0.5 M NaCl in Tris-HCl, pH 7.4. SDS-PAGE of the eluate revealed protein bands with molecular weights 78, 57, 40, 30, 16, and 12 kD. Among these, Western blotting detected myeloperoxidase (57, 40, and 12 kD) and lactoferrin (78 kD). Also, the 30-kD component had a sequence (1)I-(2)I/V-(3)G-(4)G-(5)R/H at the N-terminus that is likely to indicate the presence of neutrophilic elastase, cathepsin G, proteinase 3, and azurocidin (CAP 37) - all from the family of serprocidins. Mass spectrometry of tryptic fragments indicated the presence of the 16-kD eosinophilic cationic protein (seven peptides), 27-kD cathepsin G (eleven peptides), 27-kD azurocidin (eight peptides), 29-kD neutrophilic elastase (seven peptides), and 27-kD proteinase 3 (six peptides). Myeloperoxidase was represented by 57-, 40-, and 12-kD fragments (thirteen, ten, and four peptides, respectively). Thus, interaction with Cp of five cationic proteins, i.e. of eosinophilic cationic protein, cathepsin G, neutrophilic elastase, proteinase 3, and azurocidin is reported for the first time.
ISSN:0006-2979
1608-3040
0320-9725
DOI:10.1134/S0006297907080093