Identification of a novel eosinophil chemotactic cytokine (ECF-L) as a chitinase family protein.
A novel eosinophil chemotactic cytokine (ECF-L) was purified from the culture supernatant of splenocytes of mice by a combination of anion-exchange chromatography, Procion red-agarose affinity chromatography, size exclusion high performance liquid chromatography (HPLC), and reverse phase HPLC. The NH(2)-terminal amino acid sequence was determined by direct protein sequencing. An ECF-L cDNA clone of 1,506 nucleotides was isolated from a cDNA library, and the nucleotide sequence predicted a mature protein of 397 amino acids. A recombinant ECF-L showed a level of eosinophil chemotactic activity comparable with that of natural ECF-L, and the activity was inhibited by a monoclonal antibody to ECF-L. ECF-L also attracted T lymphocytes and bone marrow polymorphonuclear leukocytes in vitro, whereas it caused selective extravasation of eosinophils in vivo. ECF-L mRNA was highly expressed in spleen, bone marrow, lung, and heart. A comprehensive GenBank data base search revealed that ECF-L is a chitinase family protein. ECF-L retains those amino acids highly conserved among chitinase family proteins, but Asp and Glu residues essential for the proton donation in hydrolysis were replaced by Asn and Gln, respectively. Although ECF-L contains a consensus CXC sequence near the NH(2) terminus akin to chemokine family proteins, the rest of ECF-L shows poor homology with chemokines.
Owhashi
M
M
Faculty of Integrated Arts and Sciences, Tokushima University, Tokushima 770-8502, Japan. ohashi@ias.tokushima-u.ac.jp
Arita
H
H
Hayai
N
N
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D002590:Cestode Infections / Q000097:blood / Q000503:physiopathology*; D019742:Chemokines, CC / Q000737:chemistry; D019743:Chemokines, CXC / Q000737:chemistry; D002631:Chemotactic Factors, Eosinophil / Q000737:chemistry / Q000235:genetics / Q000502:physiology*; D002688:Chitinases / Q000737:chemistry / Q000235:genetics / Q000502:physiology*; D002851:Chromatography, High Pressure Liquid; D004804:Eosinophils / Q000469:parasitology / Q000502:physiology*; D005260:Female; D005786:Gene Expression Regulation; D006801:Humans; D008214:Lymphocytes / Q000737:chemistry / Q000276:immunology; D008264:Macrophages / Q000469:parasitology / Q000502:physiology; D008645:Mesocestoides; D051379:Mice; D008810:Mice, Inbred C57BL; D008815:Mice, Inbred Strains; D008969:Molecular Sequence Data; D009504:Neutrophils / Q000469:parasitology / Q000502:physiology*; D012333:RNA, Messenger / Q000235:genetics; D012549:Schistosoma japonicum; D012554:Schistosomiasis japonica / Q000097:blood / Q000503:physiopathology*; D016415:Sequence Alignment; D017386:Sequence Homology, Amino Acid; D013154:Spleen / Q000276:immunology; D014158:Transcription, Genetic
D019742:Chemokines, CC; D019743:Chemokines, CXC; D002631:Chemotactic Factors, Eosinophil; D012333:RNA, Messenger; D002688:Chitinases