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. 1973 Apr;70(4):973–976. doi: 10.1073/pnas.70.4.973

Isolation and Characterization of Two Glycoproteins from Patients with Alveolar Proteinosis

Michael A Passero 1,2,*, Ross W Tye 1,2, Kaye H Kilburn 1,2, William S Lynn 1,2
PMCID: PMC433404  PMID: 4123934

Abstract

The saline-insoluble particulate material obtained by pulmonary lavage from four patients with alveolar proteinosis was analyzed for total lipid (52%), protein (44%), and carbohydrate (4%). Sodium dodecyl sulfate-gel electrophoresis revealed only three major peptides in these patients. Molecular weights (from the gels) were 69,000, 62,000, and 36,000, and the latter two peptides were periodic acid-Schiff stain-positive. Amino-acid and sugar analyses were performed on the peptides cut from the gels and on peptides purified by Sephadex G-200 chromatography. The 69,000-molecular-weight peptide contained no carbohydrate. Antibody studies and aminoacid analysis indicated that it was albumin. The 62,000-molecular-weight peptide contained 1% hydroxyproline, 1% hydroxylysine, 10% glycine, and 9% carbohydrate. The 36,000-molecular-weight peptide contained 1.2% hydroxyproline, 1% hydroxylysine, 13% glycine, 1.4% sialic acid, 2.6% glucose, 2.4% galactose, 2% mannose, 0.8% fucose, 0.4% glycosamine, and 0.6% galactosamine. Proteins extracted from kidney glomeruli with similar amino-acid and carbohydrate composition have been observed previously.

Keywords: pulmonary lavage, hydroxyproline, hydroxylysine, alveolar glycoproteins

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Calcott M. A., Müller-Eberhard H. J. C1q protein of human complement. Biochemistry. 1972 Aug 29;11(18):3443–3450. doi: 10.1021/bi00768a018. [DOI] [PubMed] [Google Scholar]
  2. Evans W. H., Gurd J. W. Preparation and properties of nexuses and lipid-enriched vesicles from mouse liver plasma membranes. Biochem J. 1972 Jul;128(3):691–700. doi: 10.1042/bj1280691. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Guidotti G. Membrane proteins. Annu Rev Biochem. 1972;41:731–752. doi: 10.1146/annurev.bi.41.070172.003503. [DOI] [PubMed] [Google Scholar]
  4. Hawkins J. E., Savard E. V., Ramirez-Rivera J. Pulmonary alveolar proteinosis. Origin of proteins in pulmonary washings. Am J Clin Pathol. 1967 Jul;48(1):14–17. doi: 10.1093/ajcp/48.1.14. [DOI] [PubMed] [Google Scholar]
  5. Houston L. L. Amino acid analysis of stained bands from polyacrylamide gels. Anal Biochem. 1971 Nov;44(1):81–88. doi: 10.1016/0003-2697(71)90348-4. [DOI] [PubMed] [Google Scholar]
  6. Hudson B. G., Spiro R. G. Fractionation of glycoprotein components of the reduced alkylated renal glomerular basement membrane. J Biol Chem. 1972 Jul 10;247(13):4239–4247. [PubMed] [Google Scholar]
  7. Hudson B. G., Spiro R. G. Studies on the native and reduced alkylated renal glomerular basement membrane. Solubility, subunit size, and reaction with cyanogen bromide. J Biol Chem. 1972 Jul 10;247(13):4229–4238. [PubMed] [Google Scholar]
  8. Kylstra J. A., Rausch D. C., Hall K. D., Spock A. Volume-controlled lung lavage in the treatment of asthma, bronchiectasis, and mucoviscidosis. Am Rev Respir Dis. 1971 May;103(5):651–665. doi: 10.1164/arrd.1971.103.5.651. [DOI] [PubMed] [Google Scholar]
  9. LOWRY O. H., ROSEBROUGH N. J., FARR A. L., RANDALL R. J. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951 Nov;193(1):265–275. [PubMed] [Google Scholar]
  10. Lee Y. C., Johnson G. S., White B., Scocca J. An accelerated system for analysis of neutral sugars in complex carbohydrates. Anal Biochem. 1971 Oct;43(2):640–643. doi: 10.1016/0003-2697(71)90301-0. [DOI] [PubMed] [Google Scholar]
  11. Marchesi V. T., Andrews E. P. Glycoproteins: isolation from cellmembranes with lithium diiodosalicylate. Science. 1971 Dec 17;174(4015):1247–1248. doi: 10.1126/science.174.4015.1247. [DOI] [PubMed] [Google Scholar]
  12. Nachman R. L., Ferris B., Hirsch J. G. Macrophage plasma membranes. I. Isolation and studies on protein components. J Exp Med. 1971 Apr 1;133(4):785–806. doi: 10.1084/jem.133.4.785. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. PIEZ K. A., MORRIS L. A modified procedure for the automatic analysis of amino acids. Anal Biochem. 1960 Nov;1:187–201. doi: 10.1016/0003-2697(60)90045-2. [DOI] [PubMed] [Google Scholar]
  14. ROSEN S. H., CASTLEMAN B., LIEBOW A. A. Pulmonary alveolar proteinosis. N Engl J Med. 1958 Jun 5;258(23):1123–1142. doi: 10.1056/NEJM195806052582301. [DOI] [PubMed] [Google Scholar]
  15. Ramirez J., Harlan W. R., Jr Pulmonary alveolar proteinosis. Nature and origin of alveolar lipid. Am J Med. 1968 Oct;45(4):502–512. doi: 10.1016/0002-9343(68)90166-6. [DOI] [PubMed] [Google Scholar]
  16. Ramirez J. Pulmonary alveolar proteinosis. Treatment by massive bronchopulmonary lavage. Arch Intern Med. 1967 Feb;119(2):147–156. doi: 10.1001/archinte.119.2.147. [DOI] [PubMed] [Google Scholar]
  17. Spiro R. G. Studies on the renal glomerular basement membrane. Preparation and chemical composition. J Biol Chem. 1967 Apr 25;242(8):1915–1922. [PubMed] [Google Scholar]
  18. Vracko R. Significance of basal lamina for regeneration of injured lung. Virchows Arch A Pathol Pathol Anat. 1972;355(3):264–274. doi: 10.1007/BF00551062. [DOI] [PubMed] [Google Scholar]
  19. WARREN L. The thiobarbituric acid assay of sialic acids. J Biol Chem. 1959 Aug;234(8):1971–1975. [PubMed] [Google Scholar]
  20. Weber K., Osborn M. The reliability of molecular weight determinations by dodecyl sulfate-polyacrylamide gel electrophoresis. J Biol Chem. 1969 Aug 25;244(16):4406–4412. [PubMed] [Google Scholar]
  21. Yonemasu K., Stroud R. M. Structural studies on human Clq: non-covalent and covalent subunits. Immunochemistry. 1972 May;9(5):545–554. doi: 10.1016/0019-2791(72)90064-x. [DOI] [PubMed] [Google Scholar]
  22. Zacharius R. M., Zell T. E., Morrison J. H., Woodlock J. J. Glycoprotein staining following electrophoresis on acrylamide gels. Anal Biochem. 1969 Jul;30(1):148–152. doi: 10.1016/0003-2697(69)90383-2. [DOI] [PubMed] [Google Scholar]

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