Characteristics of human surfactant-associated glycoproteins A.
Surfactant-associated glycoprotein A [molecular weight (Mr) = 34,000, isoelectric point (pI) 4.6-5.0] and its sulfhydryl dependent oligomers were purified and partially characterized from surfactant obtained from human alveolar lavage. Two major forms of the protein were identified by silver stain and immunoblot analysis of surfactant using human surfactant-associated glycoprotein A antisera: glycoprotein A2, Mr = 34,000 and glycoprotein A1, Mr = 28,000. The larger form was reduced to Mr = 28,000 by treatment with endoglycosidase F, indicating the presence of complex N-linked oligosaccharide on the molecule. Charge heterogeneity was decreased and the isoelectric point increased by treatment with neuroaminidase, supporting the presence of sialic acid. Homology between the proteins Mr = 34,000 and 28,000 was confirmed by analysis of two-dimensional tryptic and chymotryptic peptides of 125I-iodo-glycoproteins A1 and A2 which were identical. The protein was very rich in glycine and its amino acid composition was similar to that of glycoprotein A previously reported for the dog and rat. Treatment of glycoproteins A with bacterial collagenase resulted in the generation of highly glycosylated peptides Mr = 20,000-22,000, pI 4.6-5.0, which no longer formed sulfhydryl-dependent oligomers, supporting the presence of significant collagen-like region in the molecule. In the absence of reducing agents, glycoprotein A from surfactant was present as sulfhydryl-dependent dimers and larger oligomers. Higher molecular weight aggregates of glycoproteins A were also present in lavage material even after sulfhydryl reduction. Glycoproteins A were identified in surfactant from amniotic fluid, normal adult lung lavage, human cadaver lung lavage, and material obtained from lung lavage from a patient with alveolar proteinosis.(ABSTRACT TRUNCATED AT 250 WORDS)
D000596:Amino Acids / Q000032:analysis; D004591:Electrophoresis, Polyacrylamide Gel; D006023:Glycoproteins / Q000032:analysis*; D006801:Humans; D007123:Immunoelectrophoresis, Two-Dimensional; D007525:Isoelectric Focusing; D010455:Peptides / Q000032:analysis; D011510:Proteolipids / Q000032:analysis*; D037661:Pulmonary Surfactant-Associated Proteins; D011663:Pulmonary Surfactants / Q000032:analysis*
D000596:Amino Acids; D006023:Glycoproteins; D010455:Peptides; D011510:Proteolipids; D037661:Pulmonary Surfactant-Associated Proteins; D011663:Pulmonary Surfactants