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PMID: 11485569 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Differential effects of glycosphingolipids on the detergent-insolubility of the glycosylphosphatidylinositol-anchored membrane dipeptidase.

The Biochemical journal ·Vol. 358 ·No. Pt 1 ·2001-08-15 ·Pages 209-16

Parkin ET, Turner AJ, Hooper NM

Abstract

The insolubility of glycosylphosphatidylinositol (GPI)-anchored proteins in certain detergents appears to be an intrinsic property of their association with sphingolipids and cholesterol in lipid rafts. We show that the GPI-anchored protein membrane dipeptidase is localized in detergent-insoluble lipid rafts isolated from porcine kidney microvillar membranes, and that these rafts, which lack caveolin, are enriched not only in sphingomyelin and cholesterol, but also in the glycosphingolipid lactosylceramide (LacCer). Dipeptidase purified from porcine kidney was reconstituted into artificial liposomes in order to investigate the relationship between glycosphingolipids and GPI-anchored protein detergent-insolubility. Dipeptidase was insoluble in liposomes containing extremely low concentrations of LacCer. In contrast, identical concentrations of glucosylceramide or galactosylceramide failed to promote significant detergent-insolubility. Cholesterol was shown to enhance the detergent-insoluble effect of LacCer. GC-MS analysis revealed dramatic differences between the fatty acyl compositions of LacCer and those of the other glycosphingolipids. However, despite these differences, we show that the unusually marked effect of LacCer to promote the detergent-insolubility of dipeptidase cannot be singularly attributed to the fatty acyl composition of this glycosphingolipid molecule. Instead, we suggest that the ability of LacCer to confer detergent-insolubility on this GPI-anchored protein is dependent on the structure of the lipid molecule in its entirety, and that this glycosphingolipid may have an important role to play in the stabilization of lipid rafts, particularly the caveolin-free glycosphingolipid signalling domains.

MeSH Terms
Animals Brain/metabolism Caveolin 1 Caveolins/chemistry Cell Membrane/enzymology Cholesterol/chemistry Detergents/pharmacology Dipeptidases/chemistry Eggs Electrophoresis, Polyacrylamide Gel Fatty Acids/chemistry,metabolism Galactosylceramides/chemistry Gas Chromatography-Mass Spectrometry Glucosylceramides/chemistry Glycosphingolipids/chemistry Immunoblotting Kidney/chemistry,enzymology,metabolism Lipids/chemistry Membrane Microdomains/chemistry,enzymology Microvilli/chemistry Protein Structure, Tertiary Signal Transduction Sphingolipids/chemistry Sphingomyelins/chemistry Swine
Chemicals
Caveolin 1 Caveolins Detergents Fatty Acids Galactosylceramides Glucosylceramides Glycosphingolipids Lipids Sphingolipids Sphingomyelins Cholesterol Dipeptidases dipeptidase
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Parkin E T
School of Biochemistry and Molecular Biology, University of Leeds, Leeds LS2 9JT, UK. [email protected]
Turner A J
Hooper N M
References (51)
51 references, click to expand
  1. Triton X-100-resistant membrane complexes from cultured kidney epithelial cells contain the Src family protein tyrosine kinase p62yes.
    J Biol Chem. 1994 Jul 22;269(29):19123-7 PMID: 7518450
  2. Lipid sorting in epithelial cells.
    Biochemistry. 1988 Aug 23;27(17):6197-202 PMID: 3064805
  3. A rapid method for the preparation of microvilli from rabbit kidney.
    Biochem J. 1974 Sep;142(3):575-81 PMID: 4464842
  4. Analysis of brain lipids by high performance thin-layer chromatography and densitometry.
    J Lipid Res. 1983 Sep;24(9):1243-50 PMID: 6631248
  5. Reconstitution of membrane proteins. Spontaneous association of integral membrane proteins with preformed unilamellar lipid bilayers.
    Biochemistry. 1985 Jul 16;24(15):4066-75 PMID: 2996590
  6. Isolation of two differentially glycosylated forms of peptidyl-dipeptidase A (angiotensin converting enzyme) from pig brain: a re-evaluation of their role in neuropeptide metabolism.
    Biochem J. 1987 Feb 1;241(3):625-33 PMID: 2439065
  7. Renal dipeptidase is one of the membrane proteins released by phosphatidylinositol-specific phospholipase C.
    Biochem J. 1987 Jun 1;244(2):465-9 PMID: 2822007
  8. Pig kidney angiotensin converting enzyme. Purification and characterization of amphipathic and hydrophilic forms of the enzyme establishes C-terminal anchorage to the plasma membrane.
    Biochem J. 1987 Oct 1;247(1):85-93 PMID: 2825659
  9. Ectoenzymes of the kidney microvillar membrane. Aminopeptidase P is anchored by a glycosyl-phosphatidylinositol moiety.
    FEBS Lett. 1988 Mar 14;229(2):340-4 PMID: 3278935
  10. Ectoenzymes of the kidney microvillar membrane. Differential solubilization by detergents can predict a glycosyl-phosphatidylinositol membrane anchor.
    Biochem J. 1988 Mar 15;250(3):865-9 PMID: 2839148
  11. Ectoenzymes of the kidney microvillar membrane. Affinity purification, characterization and localization of the phospholipase C-solubilized form of renal dipeptidase.
    Biochem J. 1989 Jan 15;257(2):361-7 PMID: 2930455
  12. Ectoenzymes of the kidney microvillar membrane. Isolation and characterization of the amphipathic form of renal dipeptidase and hydrolysis of its glycosyl-phosphatidylinositol anchor by an activity in plasma.
    Biochem J. 1989 Aug 1;261(3):811-8 PMID: 2529850
  13. Glycosphingolipids are not essential for formation of detergent-resistant membrane rafts in melanoma cells. methyl-beta-cyclodextrin does not affect cell surface transport of a GPI-anchored protein.
    J Biol Chem. 1999 Nov 26;274(48):34459-66 PMID: 10567427
  14. Acyl and alkyl chain length of GPI-anchors is critical for raft association in vitro.
    FEBS Lett. 1999 Nov 26;462(1-2):47-50 PMID: 10580089
  15. Aminopeptidase N/CD13 is associated with raft membrane microdomains in monocytes.
    Biochem Biophys Res Commun. 2000 Mar 5;269(1):143-8 PMID: 10694491
  16. Effect of synthetic sialyl 2-->1 sphingosine and other glycosylsphingosines on the structure and function of the "glycosphingolipid signaling domain (GSD)" in mouse melanoma B16 cells.
    Biochemistry. 2000 Mar 14;39(10):2459-68 PMID: 10704195
  17. Structure and function of sphingolipid- and cholesterol-rich membrane rafts.
    J Biol Chem. 2000 Jun 9;275(23):17221-4 PMID: 10770957
  18. Caveolae/lipid rafts in fibroblast-like synoviocytes: ectopeptidase-rich membrane microdomains.
    Biochem J. 2001 Feb 15;354(Pt 1):47-55 PMID: 11171078
  19. Lipid rafts and signal transduction.
    Nat Rev Mol Cell Biol. 2000 Oct;1(1):31-9 PMID: 11413487
  20. Sorting of GPI-anchored proteins to glycolipid-enriched membrane subdomains during transport to the apical cell surface.
    Cell. 1992 Feb 7;68(3):533-44 PMID: 1531449
  21. Characterization of a secretase activity which releases angiotensin-converting enzyme from the membrane.
    Biochem J. 1993 Jun 1;292 ( Pt 2):597-603 PMID: 8389141
  22. Glycosphingolipid-enriched, detergent-insoluble complexes in protein sorting in epithelial cells.
    Biochemistry. 1993 Jun 29;32(25):6365-73 PMID: 8518282
  23. Signal transducing molecules and glycosyl-phosphatidylinositol-linked proteins form a caveolin-rich insoluble complex in MDCK cells.
    J Cell Biol. 1993 Aug;122(4):789-807 PMID: 8349730
  24. VIP21/caveolin, glycosphingolipid clusters and the sorting of glycosylphosphatidylinositol-anchored proteins in epithelial cells.
    EMBO J. 1994 Jan 1;13(1):42-53 PMID: 8306971
  25. Purification and characterization of smooth muscle cell caveolae.
    J Cell Biol. 1994 Jul;126(1):127-38 PMID: 8027172
  26. Interactions between saturated acyl chains confer detergent resistance on lipids and glycosylphosphatidylinositol (GPI)-anchored proteins: GPI-anchored proteins in liposomes and cells show similar behavior.
    Proc Natl Acad Sci U S A. 1994 Dec 6;91(25):12130-4 PMID: 7991596
  27. Involvement of detergent-insoluble complexes in the intracellular transport of intestinal brush border enzymes.
    Biochemistry. 1995 Feb 7;34(5):1596-605 PMID: 7849019
  28. Both sphingolipids and cholesterol participate in the detergent insolubility of alkaline phosphatase, a glycosylphosphatidylinositol-anchored protein, in mammalian membranes.
    J Biol Chem. 1995 Mar 17;270(11):6254-60 PMID: 7890763
  29. Cholesterol depletion and modification of COOH-terminal targeting sequence of the prion protein inhibit formation of the scrapie isoform.
    J Cell Biol. 1995 Apr;129(1):121-32 PMID: 7698979
  30. Glycolipid-anchored proteins in neuroblastoma cells form detergent-resistant complexes without caveolin.
    J Cell Biol. 1995 May;129(3):619-27 PMID: 7537273
  31. Structures of the glycosyl-phosphatidylinositol anchors of porcine and human renal membrane dipeptidase. Comprehensive structural studies on the porcine anchor and interspecies comparison of the glycan core structures.
    J Biol Chem. 1995 Sep 29;270(39):22946-56 PMID: 7559431
  32. Isolation and characterization of two distinct low-density, Triton-insoluble, complexes from porcine lung membranes.
    Biochem J. 1996 Nov 1;319 ( Pt 3):887-96 PMID: 8920995
  33. Tyrosine kinase receptors concentrated in caveolae-like domains from neuronal plasma membrane.
    J Biol Chem. 1997 Feb 7;272(6):3554-9 PMID: 9013605
  34. Proteolytic fragmentation reveals the oligomeric and domain structure of porcine aminopeptidase A.
    Biochemistry. 1997 Mar 11;36(10):3000-7 PMID: 9062131
  35. Flotillin and epidermal surface antigen define a new family of caveolae-associated integral membrane proteins.
    J Biol Chem. 1997 May 23;272(21):13793-802 PMID: 9153235
  36. Functional rafts in cell membranes.
    Nature. 1997 Jun 5;387(6633):569-72 PMID: 9177342
  37. Signal transduction via glycosyl phosphatidylinositol-anchored proteins in T cells is inhibited by lowering cellular cholesterol.
    J Biol Chem. 1997 Aug 1;272(31):19242-7 PMID: 9235917
  38. On the origin of sphingolipid/cholesterol-rich detergent-insoluble cell membranes: physiological concentrations of cholesterol and sphingolipid induce formation of a detergent-insoluble, liquid-ordered lipid phase in model membranes.
    Biochemistry. 1997 Sep 9;36(36):10944-53 PMID: 9283086
  39. The amyloid precursor protein is not enriched in caveolae-like, detergent-insoluble membrane microdomains.
    J Neurochem. 1997 Nov;69(5):2179-88 PMID: 9349565
  40. Sphingolipid organization in biomembranes: what physical studies of model membranes reveal.
    J Cell Sci. 1998 Jan;111 ( Pt 1):1-9 PMID: 9394007
  41. Cholesterol and sphingolipid enhance the Triton X-100 insolubility of glycosylphosphatidylinositol-anchored proteins by promoting the formation of detergent-insoluble ordered membrane domains.
    J Biol Chem. 1998 Jan 9;273(2):1150-7 PMID: 9422781
  42. Functional organization of the Golgi apparatus in glycosphingolipid biosynthesis. Lactosylceramide and subsequent glycosphingolipids are formed in the lumen of the late Golgi.
    J Biol Chem. 1998 Jan 30;273(5):2939-46 PMID: 9446606
  43. Molecular forces between membranes displaying neutral glycosphingolipids: evidence for carbohydrate attraction.
    Biochemistry. 1998 Feb 10;37(6):1540-50 PMID: 9484224
  44. Structure and origin of ordered lipid domains in biological membranes.
    J Membr Biol. 1998 Jul 15;164(2):103-14 PMID: 9662555
  45. Glucosylceramide synthase and glycosphingolipid synthesis.
    Trends Cell Biol. 1998 May;8(5):198-202 PMID: 9695839
  46. Separation of "glycosphingolipid signaling domain" from caveolin-containing membrane fraction in mouse melanoma B16 cells and its role in cell adhesion coupled with signaling.
    J Biol Chem. 1998 Dec 11;273(50):33766-73 PMID: 9837965
  47. Functions of lipid rafts in biological membranes.
    Annu Rev Cell Dev Biol. 1998;14:111-36 PMID: 9891780
  48. Characterization of detergent-insoluble complexes containing the familial Alzheimer's disease-associated presenilins.
    J Neurochem. 1999 Apr;72(4):1534-43 PMID: 10098859
  49. Detergent-insoluble glycosphingolipid/cholesterol-rich membrane domains, lipid rafts and caveolae (review).
    Mol Membr Biol. 1999 Apr-Jun;16(2):145-56 PMID: 10417979
  50. Exogenous administration of gangliosides displaces GPI-anchored proteins from lipid microdomains in living cells.
    Mol Biol Cell. 1999 Oct;10(10):3187-96 PMID: 10512859
  51. Detergent-insoluble glycolipid microdomains in lymphocytes in the absence of caveolae.
    J Biol Chem. 1994 Dec 9;269(49):30745-8 PMID: 7982998
Article Info
Journal
The Biochemical journal
Abbr.
Biochem J
ISSN
0264-6021
DOI
10.1042/0264-6021:3580209
Email
[email protected]
Published
2001-08-15
Pages
209-16
Language
English
Region
England
NLM ID
2984726R
PMCID
PMC1222049
Subset
IM
External Links
PubMed source DOI: 10.1042/0264-6021:3580209 PMC Full Text View journal details
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