title: Epitope mapping for monoclonal antibody against human surfactant protein A (SP-A) that alters receptor binding of SP-A and the SP-A-dependent regulation of phospholipid secretion by alveolar type II cells.
abstract: Surfactant protein A (SP-A) is a lung-specific glycoprotein in pulmonary surfactant and has a collagen like sequence on its N-terminal. SP-A has been shown to function as an inhibitor of phospholipid secretion by primary culture of alveolar type II cells via cell surface receptor(s) for SP-A. In a previous report, we showed that the C-terminal non-collagen like domain of human SP-A possessed the biological activities, and that a monoclonal antibody against human SP-A, PE10, abolished the biological activity of SP-A (Murata et al. (1993) Biochem. J. 291, 71-76). In the present study, we investigated an epitope of SP-A for PE10. Western blot analysis with fragmented peptides of human SP-A generated by both lysyl endopeptidase and BrCN showed that PE10 reacted with the peptide corresponding with Glu202 to the C-terminal but that it lacked the ability to bind to the peptide corresponding with Tyr208 to the C-terminal. The antibodies against a synthetic peptide (P1) corresponding with Glu202 to Asn217 of human SP-A inhibited the binding of PE10 to SP-A, suggesting that a similar site was recognized by both PE10 and anti-P1 antibodies. Anti-P1 antibodies as well as PE10 suppressed the biological activity of SP-A. A direct interaction between P1 and rat lung membranes, or between P1 and alveolar type II cell membranes was shown from the measurement of the fluorescence emission spectra of dansyl-labeled P1. These results suggest that an area contiguous to or near the region from Glu202 to Met207 of SP-A is important for expressing the biological activities.
authors.affiliation: Department of Biochemistry, Sapporo Medical University School of Medicine, Japan.
authors.forename: N
authors.initials: N
authors.lastname: Hiraike
authors.forename: H
authors.initials: H
authors.lastname: Sohma
authors.forename: Y
authors.initials: Y
authors.lastname: Kuroki
authors.forename: T
authors.initials: T
authors.lastname: Akino
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology; D001667:Binding, Competitive; D015153:Blotting, Western; D002462:Cell Membrane / Q000378:metabolism; D002478:Cells, Cultured; D003619:Dansyl Compounds; D018604:Epitope Mapping; D006023:Glycoproteins / Q000737:chemistry* / Q000276:immunology / Q000378:metabolism; D006801:Humans; D008168:Lung / Q000378:metabolism; D008969:Molecular Sequence Data; D010446:Peptide Fragments / Q000737:chemistry / Q000276:immunology / Q000378:metabolism; D010713:Phosphatidylcholines / Q000037:antagonists & inhibitors / Q000378:metabolism*; D011510:Proteolipids / Q000737:chemistry* / Q000276:immunology / Q000494:pharmacology / Q000502:physiology; D011650:Pulmonary Alveoli / Q000166:cytology / Q000378:metabolism*; D037662:Pulmonary Surfactant-Associated Protein A; D037661:Pulmonary Surfactant-Associated Proteins; D011663:Pulmonary Surfactants / Q000737:chemistry* / Q000276:immunology / Q000494:pharmacology / Q000502:physiology; D051381:Rats; D017207:Rats, Sprague-Dawley; D012697:Serine Endopeptidases / Q000378:metabolism; D013050:Spectrometry, Fluorescence
chemical_list: D000911:Antibodies, Monoclonal; D003619:Dansyl Compounds; D006023:Glycoproteins; D010446:Peptide Fragments; D010713:Phosphatidylcholines; D011510:Proteolipids; D037662:Pulmonary Surfactant-Associated Protein A; D037661:Pulmonary Surfactant-Associated Proteins; D011663:Pulmonary Surfactants; D012697:Serine Endopeptidases; C036813:lysyl endopeptidase