Glycolipid-anchored Proteins in Neuroblastoma Cells Form Detergent-resistant Complexes Without Caveolin

Overview

Journal J Cell Biol

Specialty Cell Biology

Date 1995 May 1

PMID 7537273

Citations 89

Authors

A Gorodinsky

D A Harris

Affiliations

Soon will be listed here.

Abstract

It has been known for a number of years that glycosyl-phosphatidylinositol (GPI)-anchored proteins, in contrast to many transmembrane proteins, are insoluble at 4 degrees C in nonionic detergents such as Triton X-100. Recently, it has been proposed that this behavior reflects the incorporation of GPI-linked proteins into large aggregates that are rich in sphingolipids and cholesterol, as well as in cytoplasmic signaling molecules such as heterotrimeric G proteins and src-family tyrosine kinases. It has been suggested that these lipid-protein complexes are derived from caveolae, non-clathrin-coated invaginations of the plasmalemma that are abundant in endothelial cells, smooth muscle, and lung. Caveolin, a proposed coat protein of caveolae, has been hypothesized to be essential for formation of the complexes. To further investigate the relationship between the detergent-resistant complexes and caveolae, we have characterized the behavior of GPI-anchored proteins in lysates of N2a neuroblastoma cells, which lack morphologically identifiable caveolae, and which do not express caveolin (Shyng, S.-L., J. E. Heuser, and D. A. Harris. 1994. J. Cell Biol. 125:1239-1250). We report here that the complexes prepared from N2a cells display the large size and low buoyant density characteristic of complexes isolated from sources that are rich in caveolae, and contain the same major constituents, including multiple GPI-anchored proteins, alpha and beta subunits of heterotrimeric G proteins, and the tyrosine kinases fyn and yes. Our results argue strongly that detergent-resistant complexes are not equivalent to caveolae in all cell types, and that in neuronal cells caveolin is not essential for the integrity of these complexes.

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References

Kurzchalia T, Dupree P, Parton R, Kellner R, Virta H, Lehnert M . VIP21, a 21-kD membrane protein is an integral component of trans-Golgi-network-derived transport vesicles. J Cell Biol. 1992; 118(5):1003-14. PMC: 2289580. DOI: 10.1083/jcb.118.5.1003. View

Englund P . The structure and biosynthesis of glycosyl phosphatidylinositol protein anchors. Annu Rev Biochem. 1993; 62:121-38. DOI: 10.1146/annurev.bi.62.070193.001005. View

Cinek T, Horejsi V . The nature of large noncovalent complexes containing glycosyl-phosphatidylinositol-anchored membrane glycoproteins and protein tyrosine kinases. J Immunol. 1992; 149(7):2262-70. View

Cullen B . Trans-activation of human immunodeficiency virus occurs via a bimodal mechanism. Cell. 1986; 46(7):973-82. DOI: 10.1016/0092-8674(86)90696-3. View

Lisanti M, Tang Z, Sargiacomo M . Caveolin forms a hetero-oligomeric protein complex that interacts with an apical GPI-linked protein: implications for the biogenesis of caveolae. J Cell Biol. 1993; 123(3):595-604. PMC: 2200116. DOI: 10.1083/jcb.123.3.595. View

Lisanti M, Scherer P, Vidugiriene J, Tang Z, Hermanowski-Vosatka A, Tu Y . Characterization of caveolin-rich membrane domains isolated from an endothelial-rich source: implications for human disease. J Cell Biol. 1994; 126(1):111-26. PMC: 2120102. DOI: 10.1083/jcb.126.1.111. View

Low M . The glycosyl-phosphatidylinositol anchor of membrane proteins. Biochim Biophys Acta. 1989; 988(3):427-54. DOI: 10.1016/0304-4157(89)90014-2. View

Shyng S, Huber M, Harris D . A prion protein cycles between the cell surface and an endocytic compartment in cultured neuroblastoma cells. J Biol Chem. 1993; 268(21):15922-8. View

Shyng S, Heuser J, Harris D . A glycolipid-anchored prion protein is endocytosed via clathrin-coated pits. J Cell Biol. 1994; 125(6):1239-50. PMC: 2290925. DOI: 10.1083/jcb.125.6.1239. View

10.

Bordier C . Phase separation of integral membrane proteins in Triton X-114 solution. J Biol Chem. 1981; 256(4):1604-7. View

11.

Mayor S, Rothberg K, Maxfield F . Sequestration of GPI-anchored proteins in caveolae triggered by cross-linking. Science. 1994; 264(5167):1948-51. DOI: 10.1126/science.7516582. View

12.

Casey P, Fong H, Simon M, GILMAN A . Gz, a guanine nucleotide-binding protein with unique biochemical properties. J Biol Chem. 1990; 265(4):2383-90. View

13.

Brown D . The tyrosine kinase connection: how GPI-anchored proteins activate T cells. Curr Opin Immunol. 1993; 5(3):349-54. DOI: 10.1016/0952-7915(93)90052-t. View

14.

Smart E, Ying Y, Conrad P, Anderson R . Caveolin moves from caveolae to the Golgi apparatus in response to cholesterol oxidation. J Cell Biol. 1994; 127(5):1185-97. PMC: 2120264. DOI: 10.1083/jcb.127.5.1185. View

15.

Rothberg K, Ying Y, Kolhouse J, Kamen B, Anderson R . The glycophospholipid-linked folate receptor internalizes folate without entering the clathrin-coated pit endocytic pathway. J Cell Biol. 1990; 110(3):637-49. PMC: 2116044. DOI: 10.1083/jcb.110.3.637. View

16.

Caughey B, Raymond G . The scrapie-associated form of PrP is made from a cell surface precursor that is both protease- and phospholipase-sensitive. J Biol Chem. 1991; 266(27):18217-23. View

17.

Harris D, Falls D, Johnson F, Fischbach G . A prion-like protein from chicken brain copurifies with an acetylcholine receptor-inducing activity. Proc Natl Acad Sci U S A. 1991; 88(17):7664-8. PMC: 52362. DOI: 10.1073/pnas.88.17.7664. View

18.

Fra A, Williamson E, Simons K, Parton R . Detergent-insoluble glycolipid microdomains in lymphocytes in the absence of caveolae. J Biol Chem. 1994; 269(49):30745-8. View

19.

Mumby S, Kahn R, Manning D, GILMAN A . Antisera of designed specificity for subunits of guanine nucleotide-binding regulatory proteins. Proc Natl Acad Sci U S A. 1986; 83(2):265-9. PMC: 322838. DOI: 10.1073/pnas.83.2.265. View

20.

Brown D, Rose J . Sorting of GPI-anchored proteins to glycolipid-enriched membrane subdomains during transport to the apical cell surface. Cell. 1992; 68(3):533-44. DOI: 10.1016/0092-8674(92)90189-j. View