Skip to main content
PMC home page
Proceedings of the National Academy of Sciences of the United States of America logoLink to Proceedings of the National Academy of Sciences of the United States of America
. 1986 Apr;83(8):2501–2505. doi: 10.1073/pnas.83.8.2501

Identification of the macrophage mannose receptor as a 175-kDa membrane protein.

T E Wileman, M R Lennartz, P D Stahl
PMCID: PMC323326  PMID: 3458213

Abstract

Mannose-lactoperoxidase, a neoglycoprotein prepared by reaction of lactoperoxidase with cyanomethyl 1-thiomannoside, bound to alveolar macrophages at 4 degrees C (Kd = 5.8 X 10(-8) M) and was rapidly internalized at 37 degrees C (K uptake = 2 X 10(-8) M). Mannose-lactoperoxidase binding and uptake were blocked by yeast mannan, and mannose-lactoperoxidase inhibited uptake of 125I-labeled mannose-BSA (bovine serum albumin). Radioiodination of cells with surface-bound mannose-lactoperoxidase was carried out in the presence of glucose and glucose oxidase. A major polypeptide (175 kDa) was radioiodinated by this procedure. Iodination of the 175-kDa polypeptide appeared to be receptor-mediated, since it was blocked by the presence of yeast mannan. Specific iodination was absent from receptor-negative cells. To demonstrate that the 175-kDa species is a ligand-binding protein, cells were iodinated by the standard lactoperoxidase method. Washed cells were then allowed to bind mannose-BSA. Receptor-ligand complexes, prepared by detergent extraction, were passed over anti-BSA IgG affinity columns. Mannose, but not mannose 6-phosphate or galactose, eluted a radioactive protein from the column that migrated with an apparent molecular mass of 175 kDa on NaDodSO4/PAGE. Detergent extracts of crude membranes prepared from macrophage-enriched whole rabbit lung were adsorbed to mannose-Sepharose; the fraction obtained by elution with mannose contained two protein components of 175 and 55 kDa. Subsequent chromatography on N-acetylglucosamine-agarose yielded a single protein of 175 kDa. The 175-kDa polypeptide was shown to bind 125I-labeled mannose-BSA in a precipitation assay. This binding could be blocked with mannan or mannose-BSA. The results indicate that the cell-surface mannose receptor is a 175-kDa protein.

Full text

PDF
2501

Images in this article

2503Image
on p.2503
2503Image
on p.2503
2504Image
on p.2504
2504Image
on p.2504

Selected References

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

  1. Bradford M. M. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem. 1976 May 7;72:248–254. doi: 10.1006/abio.1976.9999. [DOI] [PubMed] [Google Scholar]
  2. Brownell M. D., Colley K. J., Baenziger J. U. Synthesis, processing, and secretion of the core-specific lectin by rat hepatocytes and hepatoma cells. J Biol Chem. 1984 Mar 25;259(6):3925–3932. [PubMed] [Google Scholar]
  3. Harding C., Levy M. A., Stahl P. Morphological analysis of ligand uptake and processing: the role of multivesicular endosomes and CURL in receptor-ligand processing. Eur J Cell Biol. 1985 Mar;36(2):230–238. [PubMed] [Google Scholar]
  4. Kawasaki T., Etoh R., Yamashina I. Isolation and characterization of a mannan-binding protein from rabbit liver. Biochem Biophys Res Commun. 1978 Apr 14;81(3):1018–1024. doi: 10.1016/0006-291x(78)91452-3. [DOI] [PubMed] [Google Scholar]
  5. Keller R. K., Touster O. Physical and chemical properties of beta-glucuronidase from the preputial gland of the female rat. J Biol Chem. 1975 Jun 25;250(12):4765–4769. [PubMed] [Google Scholar]
  6. Kohn J., Wilchek M. A new approach (cyano-transfer) for cyanogen bromide activation of Sepharose at neutral pH, which yields activated resins, free of interfering nitrogen derivatives. Biochem Biophys Res Commun. 1982 Aug;107(3):878–884. doi: 10.1016/0006-291x(82)90604-0. [DOI] [PubMed] [Google Scholar]
  7. Kozutsumi Y., Kawasaki T., Yamashina I. Isolation and characterization of a mannan-binding protein from rabbit serum. Biochem Biophys Res Commun. 1980 Jul 31;95(2):658–664. doi: 10.1016/0006-291x(80)90836-0. [DOI] [PubMed] [Google Scholar]
  8. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  9. Leake E. S., Myrvik Q. N. Changes in morphology and in lysozyme content of free alveolar cells after the intravenous injection of killed BCG in oil. J Reticuloendothel Soc. 1968 Feb;5(1):33–53. [PubMed] [Google Scholar]
  10. Lee Y. C., Stowell C. P., Krantz M. J. 2-Imino-2-methoxyethyl 1-thioglycosides: new reagents for attaching sugars to proteins. Biochemistry. 1976 Sep 7;15(18):3956–3963. doi: 10.1021/bi00663a008. [DOI] [PubMed] [Google Scholar]
  11. Lin H. S., Gordon S. Secretion of plasminogen activator by bone marrow-derived mononuclear phagocytes and its enhancement by colony-stimulating factor. J Exp Med. 1979 Aug 1;150(2):231–245. doi: 10.1084/jem.150.2.231. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Maynard Y., Baenziger J. U. Characterization of a mannose and N-acetylglucosamine-specific lectin present in rat hepatocytes. J Biol Chem. 1982 Apr 10;257(7):3788–3794. [PubMed] [Google Scholar]
  13. Mellman I. S., Steinman R. M., Unkeless J. C., Cohn Z. A. Selective iodination and polypeptide composition of pinocytic vesicles. J Cell Biol. 1980 Sep;86(3):712–722. doi: 10.1083/jcb.86.3.712. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Shepherd V. L., Lee Y. C., Schlesinger P. H., Stahl P. D. L-Fucose-terminated glycoconjugates are recognized by pinocytosis receptors on macrophages. Proc Natl Acad Sci U S A. 1981 Feb;78(2):1019–1022. doi: 10.1073/pnas.78.2.1019. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Stahl P. D., Rodman J. S., Miller M. J., Schlesinger P. H. Evidence for receptor-mediated binding of glycoproteins, glycoconjugates, and lysosomal glycosidases by alveolar macrophages. Proc Natl Acad Sci U S A. 1978 Mar;75(3):1399–1403. doi: 10.1073/pnas.75.3.1399. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Stahl P., Schlesinger P. H., Sigardson E., Rodman J. S., Lee Y. C. Receptor-mediated pinocytosis of mannose glycoconjugates by macrophages: characterization and evidence for receptor recycling. Cell. 1980 Jan;19(1):207–215. doi: 10.1016/0092-8674(80)90402-x. [DOI] [PubMed] [Google Scholar]
  17. Steinman R. M., Mellman I. S., Muller W. A., Cohn Z. A. Endocytosis and the recycling of plasma membrane. J Cell Biol. 1983 Jan;96(1):1–27. doi: 10.1083/jcb.96.1.1. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Tietze C., Schlesinger P., Stahl P. Mannose-specific endocytosis receptor of alveolar macrophages: demonstration of two functionally distinct intracellular pools of receptor and their roles in receptor recycling. J Cell Biol. 1982 Feb;92(2):417–424. doi: 10.1083/jcb.92.2.417. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Townsend R., Stahl P. Isolation and characterization of a mannose/N-acetylglucosamine/fucose-binding protein from rat liver. Biochem J. 1981 Jan 15;194(1):209–214. doi: 10.1042/bj1940209. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Wessel D., Flügge U. I. A method for the quantitative recovery of protein in dilute solution in the presence of detergents and lipids. Anal Biochem. 1984 Apr;138(1):141–143. doi: 10.1016/0003-2697(84)90782-6. [DOI] [PubMed] [Google Scholar]
  21. Wileman T., Boshans R. L., Schlesinger P., Stahl P. Monensin inhibits recycling of macrophage mannose-glycoprotein receptors and ligand delivery to lysosomes. Biochem J. 1984 Jun 15;220(3):665–675. doi: 10.1042/bj2200665. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Wileman T., Boshans R., Stahl P. Uptake and transport of mannosylated ligands by alveolar macrophages. Studies on ATP-dependent receptor-ligand dissociation. J Biol Chem. 1985 Jun 25;260(12):7387–7393. [PubMed] [Google Scholar]

Articles from Proceedings of the National Academy of Sciences of the United States of America are provided here courtesy of National Academy of Sciences

RESOURCES