Isolation and characterization of a macrophage-derived high molecular weight protein involved in the regulation of mucus-like glycoconjugate secretion.
Pulmonary macrophages release a variety of mediators that are involved in inflammatory processes and probably are involved in respiratory mucus secretion. Conditioned media obtained from activated pulmonary macrophages were found to contain a protein that functioned as a secretagogue for mucus-like glycoconjugate (MLGC) in an in vitro bioassay. A human macrophage-derived hybridoma cell line, HB-63, exhibited the same properties and was very useful in obtaining large amounts of the protein for purification and characterization. With ultrafiltration membranes and gel electrophoresis, the protein isolated from the conditioned media of zymosan- or lipopolysaccharide-treated cells was found to have a molecular weight of approximately 68 kd. The purified protein obtained from hybridoma cells and from pulmonary macrophages exhibited strong biologic activity when it was used to stimulate MLGC secretion, both in human airway explants and in an in vitro human secretory epithelial cancer cell line. The proteins from both sources were found to have similar amino acid compositions. Preliminary results indicate the presence of the 68 kd protein in the bronchoalveolar lavage fluid of a patient with severe chronic bronchitis and mucus hypersecretion. The role of this novel protein in the lungs is, so far, speculative. The 68 kd protein may be a useful tool for studying the biosynthesis and regulation of MLGC secretion and hypersecretion.
Gollub
E G
EG
Division of Pulmonary and Critical Care Medicine, Mt. Sinai Medical Center, New York, NY 10029.
Goswami
S K
SK
Sperber
K
K
Marom
Z
Z
D000596:Amino Acids / Q000032:analysis; D000906:Antibodies / Q000302:isolation & purification; D001681:Biological Assay / Q000379:methods; D015153:Blotting, Western / Q000379:methods; D002460:Cell Line; D002469:Cell Separation; D004591:Electrophoresis, Polyacrylamide Gel / Q000379:methods; D006001:Glycoconjugates / Q000032:analysis / Q000378:metabolism*; D006023:Glycoproteins / Q000276:immunology / Q000302:isolation & purification / Q000494:pharmacology*; D006801:Humans; D006825:Hybridomas / Q000276:immunology; D015151:Immunoblotting / Q000379:methods; D016676:Macrophages, Alveolar / Q000378:metabolism*; D008970:Molecular Weight; D009093:Mucus / Q000187:drug effects* / Q000378:metabolism; D011506:Proteins / Q000032:analysis / Q000276:immunology / Q000302:isolation & purification / Q000494:pharmacology*
D000596:Amino Acids; D000906:Antibodies; D006001:Glycoconjugates; D006023:Glycoproteins; C073416:MMS-68 protein, human; D011506:Proteins