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 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.
Sokolov
A V
AV
Institute for Experimental Medicine, Russian Academy of Medical Sciences, St. Petersburg, 197376, Russia. biochem@nm.ru
Pulina
M O
MO
Ageeva
K V
KV
Runova
O L
OL
Zakharova
E T
ET
Vasil'ev
V B
VB
D002412:Cations / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism; D002570:Ceruloplasmin / Q000737:chemistry* / Q000378:metabolism; D002846:Chromatography, Affinity; D006801:Humans; D007162:Immunoproteins / Q000737:chemistry* / Q000302:isolation & purification / Q000378:metabolism; D007962:Leukocytes / Q000737:chemistry* / Q000378:metabolism; D013058:Mass Spectrometry; D011485:Protein Binding / Q000502:physiology
D002412:Cations; D007162:Immunoproteins; D002570:Ceruloplasmin