Abstract
Ligand affinity chromatography was used to purify a cell surface alpha 2-macroglobulin (alpha 2M) receptor. Detergent extracts of human placenta were applied to an affinity matrix consisting of alpha 2M, previously reacted with methylamine, coupled to Sepharose. Elution with EDTA specifically released polypeptides with apparent molecular masses of 420 and 39 kD. In some preparations, small amounts of a 90-kD polypeptide were observed. The 420- and 39-kD polypeptides appear specific for the forms of alpha 2M activated by reaction with proteinases or methylamine and do not bind to an affinity matrix consisting of native alpha 2M coupled to Sepharose. Separation of these two polypeptides was accomplished by anion exchange chromatography, and binding activity was exclusively associated with the 420-kD polypeptide. The purified 420-kD protein binds to the conformationally altered forms of alpha 2M that are known to specifically interact with alpha 2M receptors and does not bind to native alpha 2M. Binding of the 420-kD polypeptide to immobilized wheat germ agglutinin indicates that this polypeptide is a glycoprotein. The cell surface localization of the 420-kD glycoprotein was confirmed by affinity chromatography of extracts from surface radioiodinated fibroblasts. These properties suggest that the 420-kD polypeptide is a cell surface receptor for the activated forms of alpha 2M.
Full Text
The Full Text of this article is available as a PDF (1.2 MB).
Selected References
These references are in PubMed. This may not be the complete list of references from this article.
- Barak L. S., Webb W. W. Fluorescent low density lipoprotein for observation of dynamics of individual receptor complexes on cultured human fibroblasts. J Cell Biol. 1981 Sep;90(3):595–604. doi: 10.1083/jcb.90.3.595. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Barrett A. J. Alpha 2-macroglobulin. Methods Enzymol. 1981;80(Pt 100):737–754. doi: 10.1016/s0076-6879(81)80056-0. [DOI] [PubMed] [Google Scholar]
- Barrett A. J., Brown M. A., Sayers C. A. The electrophoretically 'slow' and 'fast' forms of the alpha 2-macroglobulin molecule. Biochem J. 1979 Aug 1;181(2):401–418. doi: 10.1042/bj1810401. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Björk I., Fish W. W. Evidence for similar conformational changes in alpha 2-macroglobulin on reaction with primary amines or proteolytic enzymes. Biochem J. 1982 Nov 1;207(2):347–356. doi: 10.1042/bj2070347. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Brown M. S., Anderson R. G., Goldstein J. L. Recycling receptors: the round-trip itinerary of migrant membrane proteins. Cell. 1983 Mar;32(3):663–667. doi: 10.1016/0092-8674(83)90052-1. [DOI] [PubMed] [Google Scholar]
- Cheng S. Y., Maxfield F. R., Robbins J., Willingham M. C., Pastan I. H. Receptor-mediated uptake of 3,3',5-triiodo-L-thyronine by cultured fibroblasts. Proc Natl Acad Sci U S A. 1980 Jun;77(6):3425–3429. doi: 10.1073/pnas.77.6.3425. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Dennis P. A., Saksela O., Harpel P., Rifkin D. B. Alpha 2-macroglobulin is a binding protein for basic fibroblast growth factor. J Biol Chem. 1989 May 5;264(13):7210–7216. [PubMed] [Google Scholar]
- Dickson R. B., Willingham M. C., Pastan I. Binding and internalization of 125I-alpha 2-macroglobulin by cultured fibroblasts. J Biol Chem. 1981 Apr 10;256(7):3454–3459. [PubMed] [Google Scholar]
- Dickson R. B., Willingham M. C., Pastan I. alpha 2-macroglobulin adsorbed to colloidal gold: a new probe in the study of receptor-mediated endocytosis. J Cell Biol. 1981 Apr;89(1):29–34. doi: 10.1083/jcb.89.1.29. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Feldman S. R., Rosenberg M. R., Ney K. A., Michalopoulos G., Pizzo S. V. Binding of alpha 2-macroglobulin to hepatocytes: mechanism of in vivo clearance. Biochem Biophys Res Commun. 1985 Apr 30;128(2):795–802. doi: 10.1016/0006-291x(85)90117-2. [DOI] [PubMed] [Google Scholar]
- Frey J., Afting E. G. Isolation and renaturation of alpha 2-macroglobulin receptor from diploid human fibroblasts. Biochem J. 1983 Aug 15;214(2):629–631. doi: 10.1042/bj2140629. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Gliemann J., Davidsen O. Characterization of receptors for alpha 2-macroglobulin-trypsin complex in rat hepatocytes. Biochim Biophys Acta. 1986 Jan 23;885(1):49–57. doi: 10.1016/0167-4889(86)90037-6. [DOI] [PubMed] [Google Scholar]
- Gliemann J., Davidsen O., Moestrup S. K. Characterization, size estimation and solubilization of alpha-macroglobulin complex receptors in liver membranes. Biochim Biophys Acta. 1989 Apr 28;980(3):326–332. doi: 10.1016/0005-2736(89)90320-9. [DOI] [PubMed] [Google Scholar]
- Gliemann J., Sonne O. Uptake and degradation of insulin and alpha 2-macroglobulin-trypsin complex in rat adipocytes. Evidence for different pathways. Biochim Biophys Acta. 1985 Apr 22;845(1):124–130. doi: 10.1016/0167-4889(85)90063-1. [DOI] [PubMed] [Google Scholar]
- Gonias S. L., Reynolds J. A., Pizzo S. V. Physical properties of human alpha 2-macroglobulin following reaction with methylamine and trypsin. Biochim Biophys Acta. 1982 Aug 10;705(3):306–314. doi: 10.1016/0167-4838(82)90252-7. [DOI] [PubMed] [Google Scholar]
- Hanover J. A., Cheng S., Willingham M. C., Pastan I. H. alpha 2-Macroglobulin binding to cultured fibroblasts. Solubilization and partial purification of binding sites. J Biol Chem. 1983 Jan 10;258(1):370–377. [PubMed] [Google Scholar]
- Hanover J. A., D'Souza P., August T., Pastan I., Willingham M. C. Monoclonal antibodies against a glycoprotein localized in coated pits and endocytic vesicles inhibit alpha 2-macroglobulin binding and uptake. J Biol Chem. 1986 Dec 15;261(35):16732–16737. [PubMed] [Google Scholar]
- Hanover J. A., Rudick J. E., Willingham M. C., Pastan I. Alpha 2-macroglobulin binding to cultured fibroblasts: identification by affinity chromatography of high-affinity binding sites. Arch Biochem Biophys. 1983 Dec;227(2):570–579. doi: 10.1016/0003-9861(83)90486-1. [DOI] [PubMed] [Google Scholar]
- Hanover J. A., Willingham M. C., Pastan I. Kinetics of transit of transferrin and epidermal growth factor through clathrin-coated membranes. Cell. 1984 Dec;39(2 Pt 1):283–293. doi: 10.1016/0092-8674(84)90006-0. [DOI] [PubMed] [Google Scholar]
- Herz J., Hamann U., Rogne S., Myklebost O., Gausepohl H., Stanley K. K. Surface location and high affinity for calcium of a 500-kd liver membrane protein closely related to the LDL-receptor suggest a physiological role as lipoprotein receptor. EMBO J. 1988 Dec 20;7(13):4119–4127. doi: 10.1002/j.1460-2075.1988.tb03306.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Huang J. S., Huang S. S., Deuel T. F. Specific covalent binding of platelet-derived growth factor to human plasma alpha 2-macroglobulin. Proc Natl Acad Sci U S A. 1984 Jan;81(2):342–346. doi: 10.1073/pnas.81.2.342. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Imber M. J., Pizzo S. V. Clearance and binding of two electrophoretic "fast" forms of human alpha 2-macroglobulin. J Biol Chem. 1981 Aug 10;256(15):8134–8139. [PubMed] [Google Scholar]
- Ingham K. C., Brew S. A., Isaacs B. S. Interaction of fibronectin and its gelatin-binding domains with fluorescent-labeled chains of type I collagen. J Biol Chem. 1988 Apr 5;263(10):4624–4628. [PubMed] [Google Scholar]
- Isaacs I. J., Steiner J. P., Roche P. A., Pizzo S. V., Strickland D. K. Use of anti-idiotypic antibodies to establish that monoclonal antibody 7H11D6 binds to the alpha 2-macroglobulin receptor recognition site. J Biol Chem. 1988 May 15;263(14):6709–6714. [PubMed] [Google Scholar]
- Jensen P. H., Davidsen O., Gliemann J., Petersen C. M. Cell association and degradation of pregnancy zone protein-chymotrypsin complex in cultured human monocytes. Scand J Clin Lab Invest. 1988 Apr;48(2):165–176. doi: 10.3109/00365518809085409. [DOI] [PubMed] [Google Scholar]
- Jensen P. H., Moestrup S. K., Sottrup-Jensen L., Petersen C. M., Gliemann J. Receptors for alpha 2-macroglobulin- and pregnancy zone protein-proteinase complexes in the human placental syncytiotrophoblast. Placenta. 1988 Sep-Oct;9(5):463–477. doi: 10.1016/0143-4004(88)90019-7. [DOI] [PubMed] [Google Scholar]
- Kaplan J., Keogh E. A. Analysis of the effect of amines on inhibition of receptor-mediated and fluid-phase pinocytosis in rabbit alveolar macrophages. Cell. 1981 Jun;24(3):925–932. doi: 10.1016/0092-8674(81)90118-5. [DOI] [PubMed] [Google Scholar]
- Kaplan J., Keogh E. A. Studies on the physiology of macrophage receptors for alpha-macroglobulin X protease complexes. Ann N Y Acad Sci. 1983;421:442–456. doi: 10.1111/j.1749-6632.1983.tb18138.x. [DOI] [PubMed] [Google Scholar]
- Kaplan J., Nielsen M. L. Analysis of macrophage surface receptors. I. Binding of alpha-macroglobulin . protease complexes to rabbit alveolar macrophages. J Biol Chem. 1979 Aug 10;254(15):7323–7328. [PubMed] [Google Scholar]
- Kaplan J., Ray F. A., Keogh E. A. Recognition of nucleophile-treated alpha 2-macroglobulin by the alveolar macrophage alpha-macroglobulin . protease complex receptor. J Biol Chem. 1981 Aug 10;256(15):7705–7707. [PubMed] [Google Scholar]
- Klickstein L. B., Wong W. W., Smith J. A., Weis J. H., Wilson J. G., Fearon D. T. Human C3b/C4b receptor (CR1). Demonstration of long homologous repeating domains that are composed of the short consensus repeats characteristics of C3/C4 binding proteins. J Exp Med. 1987 Apr 1;165(4):1095–1112. doi: 10.1084/jem.165.4.1095. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Marynen P., Van Leuven F., Cassiman J. J., Van den Berghe H. A monoclonal antibody to a neo-antigen on alpha 2-macroglobulin complexes inhibits receptor-mediated endocytosis. J Immunol. 1981 Nov;127(5):1782–1786. [PubMed] [Google Scholar]
- Marynen P., Van Leuven F., Cassiman J. J., Van den Berghe H. Solubilization and affinity purification of the alpha 2-macroglobulin receptor from human fibroblasts. J Biol Chem. 1984 Jun 10;259(11):7075–7079. [PubMed] [Google Scholar]
- Maxfield F. R. Weak bases and ionophores rapidly and reversibly raise the pH of endocytic vesicles in cultured mouse fibroblasts. J Cell Biol. 1982 Nov;95(2 Pt 1):676–681. doi: 10.1083/jcb.95.2.676. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Maxfield F. R., Willingham M. C., Haigler H. T., Dragsten P., Pastan I. H. Binding, surface mobility, internalization, and degradation of rhodamine-labeled alpha 2-macroglobulin. Biochemistry. 1981 Sep 1;20(18):5353–5358. doi: 10.1021/bi00521a041. [DOI] [PubMed] [Google Scholar]
- Moestrup S. K., Gliemann J. Purification of the rat hepatic alpha 2-macroglobulin receptor as an approximately 440-kDa single chain protein. J Biol Chem. 1989 Sep 15;264(26):15574–15577. [PubMed] [Google Scholar]
- Munson P. J., Rodbard D. Ligand: a versatile computerized approach for characterization of ligand-binding systems. Anal Biochem. 1980 Sep 1;107(1):220–239. doi: 10.1016/0003-2697(80)90515-1. [DOI] [PubMed] [Google Scholar]
- Ney K. A., Gidwitz S., Pizzo S. V. Changes in the binding of "fast"-form alpha 2-macroglobulin to 3T3-L1 cells after differentiation to adipocytes. Biochemistry. 1984 Jul 17;23(15):3395–3403. doi: 10.1021/bi00310a003. [DOI] [PubMed] [Google Scholar]
- O'Connor-McCourt M. D., Wakefield L. M. Latent transforming growth factor-beta in serum. A specific complex with alpha 2-macroglobulin. J Biol Chem. 1987 Oct 15;262(29):14090–14099. [PubMed] [Google Scholar]
- Petersen C. M., Christiansen B. S., Jensen P. H., Moestrup S. K., Gliemann J., Sottrup-Jensen L., Ingerslev J. Human hepatocytes exhibit receptors for alpha 2-macroglobulin and pregnancy zone protein-proteinase complexes. Eur J Clin Invest. 1988 Apr;18(2):184–190. doi: 10.1111/j.1365-2362.1988.tb02411.x. [DOI] [PubMed] [Google Scholar]
- Pytela R., Pierschbacher M. D., Argraves S., Suzuki S., Ruoslahti E. Arginine-glycine-aspartic acid adhesion receptors. Methods Enzymol. 1987;144:475–489. doi: 10.1016/0076-6879(87)44196-7. [DOI] [PubMed] [Google Scholar]
- Schneider C., Sutherland R., Newman R., Greaves M. Structural features of the cell surface receptor for transferrin that is recognized by the monoclonal antibody OKT9. J Biol Chem. 1982 Jul 25;257(14):8516–8522. [PubMed] [Google Scholar]
- Schneider W. J., Beisiegel U., Goldstein J. L., Brown M. S. Purification of the low density lipoprotein receptor, an acidic glycoprotein of 164,000 molecular weight. J Biol Chem. 1982 Mar 10;257(5):2664–2673. [PubMed] [Google Scholar]
- Siegel T. W., Ganguly S., Jacobs S., Rosen O. M., Rubin C. S. Purification and properties of the human placental insulin receptor. J Biol Chem. 1981 Sep 10;256(17):9266–9273. [PubMed] [Google Scholar]
- Sottrup-Jensen L., Stepanik T. M., Kristensen T., Lønblad P. B., Jones C. M., Wierzbicki D. M., Magnusson S., Domdey H., Wetsel R. A., Lundwall A. Common evolutionary origin of alpha 2-macroglobulin and complement components C3 and C4. Proc Natl Acad Sci U S A. 1985 Jan;82(1):9–13. doi: 10.1073/pnas.82.1.9. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Straight D. L., Jakoi L., McKee P. A., Snyderman R. Binding of alpha 2-macroglobulin-thrombin complexes and methylamine-treated alpha 2-macroglobulin to human blood monocytes. Biochemistry. 1988 Apr 19;27(8):2885–2890. doi: 10.1021/bi00408a033. [DOI] [PubMed] [Google Scholar]
- Strickland D. K., Bhattacharya P., Olson S. T. Kinetics of the conformational alterations associated with nucleophilic modification of alpha 2-macroglobulin. Biochemistry. 1984 Jul 3;23(14):3115–3124. doi: 10.1021/bi00309a002. [DOI] [PubMed] [Google Scholar]
- Strickland D. K., Steiner J. P., Migliorini M., Battey F. D. Identification of a monoclonal antibody specific for a neoantigenic determinant on alpha 2-macroglobulin: use for the purification and characterization of binary proteinase-inhibitor complexes. Biochemistry. 1988 Mar 8;27(5):1458–1466. doi: 10.1021/bi00405a010. [DOI] [PubMed] [Google Scholar]
- Turkewitz A. P., Amatruda J. F., Borhani D., Harrison S. C., Schwartz A. L. A high yield purification of the human transferrin receptor and properties of its major extracellular fragment. J Biol Chem. 1988 Jun 15;263(17):8318–8325. [PubMed] [Google Scholar]
- Van Leuven F., Cassiman J. J., Van Den Berghe H. Demonstration of an alpha2-macroglobulin receptor in human fibroblasts, absent in tumor-derived cell lines. J Biol Chem. 1979 Jun 25;254(12):5155–5160. [PubMed] [Google Scholar]
- Van Leuven F., Marynen P., Sottrup-Jensen L., Cassiman J. J., Van den Berghe H. The receptor-binding domain of human alpha 2-macroglobulin. Isolation after limited proteolysis with a bacterial proteinase. J Biol Chem. 1986 Aug 25;261(24):11369–11373. [PubMed] [Google Scholar]
- Willingham M. C., Maxfield F. R., Pastan I. H. alpha 2 Macroglobulin binding to the plasma membrane of cultured fibroblasts. Diffuse binding followed by clustering in coated regions. J Cell Biol. 1979 Sep;82(3):614–625. doi: 10.1083/jcb.82.3.614. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Willingham M. C., Maxfield F. R., Pastan I. Receptor-mediated endocytosis of alpha 2-macroglobulin in cultured fibroblasts. J Histochem Cytochem. 1980 Aug;28(8):818–823. doi: 10.1177/28.8.6160180. [DOI] [PubMed] [Google Scholar]