Journal of Cell Science, Vol 110, Issue 10 1169-1178, Copyright © 1997 by Company of Biologists

JOURNAL ARTICLES

Plasma membrane targetting, vesicular budding and release of galectin 3 from the cytoplasm of mammalian cells during secretion

B Mehul and RC Hughes
The National Institute for Medical Research, The Ridgeway, Mill Hill, London, UK.

Galectin 3, a 30 kDa galactoside-binding protein distributed widely in epithelial and immune cells, contains no signal sequence and is externalized by a mechanism independent of the endoplasmic reticulum (ER)-Golgi complex. We show here that hamster galectin 3 overexpressed in transfected cos-7 cells is secreted at a very low rate. A chimaera of galectin 3 fused to the N-terminal acylation sequence of protein tyrosine kinase p56(lck), Nt-p56(lck)-galectin 3, which is myristoylated and palmitoylated and rapidly transported to plasma membrane domains, is efficiently released from transfected cells indicating that movement of cytoplasmic galectin 3 to plasma membrane domains is a rate limiting step in lectin secretion. N-terminal acylation is not sufficient for protein secretion since p56(lck) and the chimaera Nt-p56(lck)-CAT are not secreted from transfected cells. The amino-terminal half of galectin 3 is sufficient to direct export of a chimaeric CAT protein indicating that part of the signal for plasma membrane translocation lies in the N-terminal domains of the lectin. Immunofluorescence studies show that Nt-p56(lck)-galectin 3 aggregates underneath the plasma membrane and is released by membrane blebbing. Vesicles of low buoyant density isolated from conditioned medium are enriched in galectin 3. The lectin is initially protected from exogenous collagenase but is later released in soluble protease-sensitive form from the lectin-loaded vesicles. Using murine macrophages, which secrete their endogenous galectin 3 at a moderate rate especially in the presence of Ca2+-ionophores, we were also able to trap a galectin 3-loaded vesicular fraction which was released into the culture supernatant.

This article has been cited by other articles:

				S. P. Rao, Z. Wang, R. I. Zuberi, L. Sikora, N. S. Bahaie, B. L. Zuraw, F.-T. Liu, and P. Sriramarao Galectin-3 Functions as an Adhesion Molecule to Support Eosinophil Rolling and Adhesion under Conditions of Flow J. Immunol., December 1, 2007; 179(11): 7800 - 7807. [Abstract] [Full Text] [PDF]

				E Lippert, W Falk, F Bataille, T Kaehne, M Naumann, M Goeke, H Herfarth, J Schoelmerich, and G Rogler Soluble galectin-3 is a strong, colonic epithelial-cell-derived, lamina propria fibroblast-stimulating factor Gut, January 1, 2007; 56(1): 43 - 51. [Abstract] [Full Text] [PDF]

				N. Mor-Vaknin, A. Punturieri, K. Sitwala, N. Faulkner, M. Legendre, M. S. Khodadoust, F. Kappes, J. H. Ruth, A. Koch, D. Glass, et al. The DEK Nuclear Autoantigen Is a Secreted Chemotactic Factor Mol. Cell. Biol., December 15, 2006; 26(24): 9484 - 9496. [Abstract] [Full Text] [PDF]

				M. Nagae, N. Nishi, T. Murata, T. Usui, T. Nakamura, S. Wakatsuki, and R. Kato Crystal Structure of the Galectin-9 N-terminal Carbohydrate Recognition Domain from Mus musculus Reveals the Basic Mechanism of Carbohydrate Recognition J. Biol. Chem., November 24, 2006; 281(47): 35884 - 35893. [Abstract] [Full Text] [PDF]

				C. Zehe, A. Engling, S. Wegehingel, T. Schafer, and W. Nickel Cell-surface heparan sulfate proteoglycans are essential components of the unconventional export machinery of FGF-2 PNAS, October 17, 2006; 103(42): 15479 - 15484. [Abstract] [Full Text] [PDF]

				R. Jin, A. Greenwald, M. D. Peterson, and T. K. Waddell Human Monocytes Recognize Porcine Endothelium via the Interaction of Galectin 3 and {alpha}-GAL J. Immunol., July 15, 2006; 177(2): 1289 - 1295. [Abstract] [Full Text] [PDF]

				N. C. Henderson, A. C. Mackinnon, S. L. Farnworth, F. Poirier, F. P. Russo, J. P. Iredale, C. Haslett, K. J. Simpson, and T. Sethi Galectin-3 regulates myofibroblast activation and hepatic fibrosis PNAS, March 28, 2006; 103(13): 5060 - 5065. [Abstract] [Full Text] [PDF]

				C. Seelenmeyer, S. Wegehingel, I. Tews, M. Kunzler, M. Aebi, and W. Nickel Cell surface counter receptors are essential components of the unconventional export machinery of galectin-1 J. Cell Biol., October 24, 2005; 171(2): 373 - 381. [Abstract] [Full Text] [PDF]

				D. Duzhyy, M. Harvey, and B. Sokolowski A Secretory-type Protein, Containing a Pentraxin Domain, Interacts with an A-type K+ Channel J. Biol. Chem., April 15, 2005; 280(15): 15165 - 15172. [Abstract] [Full Text] [PDF]

				R.-Y. Yang, D. K. Hsu, L. Yu, H.-Y. Chen, and F.-T. Liu Galectin-12 Is Required for Adipogenic Signaling and Adipocyte Differentiation J. Biol. Chem., July 9, 2004; 279(28): 29761 - 29766. [Abstract] [Full Text] [PDF]

				R. Backhaus, C. Zehe, S. Wegehingel, A. Kehlenbach, B. Schwappach, and W. Nickel Unconventional protein secretion: membrane translocation of FGF-2 does not require protein unfolding J. Cell Sci., May 1, 2004; 117(9): 1727 - 1736. [Abstract] [Full Text] [PDF]

				T. Schafer, H. Zentgraf, C. Zehe, B. Brugger, J. Bernhagen, and W. Nickel Unconventional Secretion of Fibroblast Growth Factor 2 Is Mediated by Direct Translocation across the Plasma Membrane of Mammalian Cells J. Biol. Chem., February 20, 2004; 279(8): 6244 - 6251. [Abstract] [Full Text] [PDF]

				M. Tanudji, S. Hevi, and S. L. Chuck The nonclassic secretion of thioredoxin is not sensitive to redox state Am J Physiol Cell Physiol, May 1, 2003; 284(5): C1272 - C1279. [Abstract] [Full Text] [PDF]

				B. K. Shin, H. Wang, A. M. Yim, F. Le Naour, F. Brichory, J. H. Jang, R. Zhao, E. Puravs, J. Tra, C. W. Michael, et al. Global Profiling of the Cell Surface Proteome of Cancer Cells Uncovers an Abundance of Proteins with Chaperone Function J. Biol. Chem., February 21, 2003; 278(9): 7607 - 7616. [Abstract] [Full Text] [PDF]

				S. G. Correa, C. E. Sotomayor, M. P. Aoki, C. A. Maldonado, and G. A. Rabinovich Opposite effects of galectin-1 on alternative metabolic pathways of L-arginine in resident, inflammatory, and activated macrophages Glycobiology, February 1, 2003; 13(2): 119 - 128. [Abstract] [Full Text] [PDF]

				A. Engling, R. Backhaus, C. Stegmayer, C. Zehe, C. Seelenmeyer, A. Kehlenbach, B. Schwappach, S. Wegehingel, and W. Nickel Biosynthetic FGF-2 is targeted to non-lipid raft microdomains following translocation to the extracellular surface of CHO cells J. Cell Sci., September 15, 2002; 115(18): 3619 - 3631. [Abstract] [Full Text] [PDF]

				G. A. Rabinovich, N. Rubinstein, and L. Fainboim Unlocking the secrets of galectins: a challenge at the frontier of glyco-immunology J. Leukoc. Biol., May 1, 2002; 71(5): 741 - 752. [Abstract] [Full Text] [PDF]

				J. L. Dunphy, G. J. Barcham, R. J. Bischof, A. R. Young, A. Nash, and E. N. T. Meeusen Isolation and Characterization of a Novel Eosinophil-specific Galectin Released into the Lungs in Response to Allergen Challenge J. Biol. Chem., April 19, 2002; 277(17): 14916 - 14924. [Abstract] [Full Text] [PDF]

				M. J. McConville, K. A. Mullin, S. C. Ilgoutz, and R. D. Teasdale Secretory Pathway of Trypanosomatid Parasites Microbiol. Mol. Biol. Rev., March 1, 2002; 66(1): 122 - 154. [Abstract] [Full Text] [PDF]

				M. Tanudji, S. Hevi, and S. L. Chuck Improperly folded green fluorescent protein is secreted via a non-classical pathway J. Cell Sci., January 10, 2002; 115(19): 3849 - 3857. [Abstract] [Full Text] [PDF]

				E. Zuniga, A. Gruppi, J. Hirabayashi, K. I. Kasai, and G. A. Rabinovich Regulated Expression and Effect of Galectin-1 on Trypanosoma cruzi-Infected Macrophages: Modulation of Microbicidal Activity and Survival Infect. Immun., November 1, 2001; 69(11): 6804 - 6812. [Abstract] [Full Text] [PDF]

				E. A. M. Barboni, S. Bawumia, K. Henrick, and R.C. Hughes Molecular modeling and mutagenesis studies of the N-terminal domains of galectin-3: evidence for participation with the C-terminal carbohydrate recognition domain in oligosaccharide binding Glycobiology, November 1, 2000; 10(11): 1201 - 1208. [Abstract] [Full Text] [PDF]

				C. Fradin, D. Poulain, and T. Jouault beta -1,2-Linked Oligomannosides from Candida albicans Bind to a 32-Kilodalton Macrophage Membrane Protein Homologous to the Mammalian Lectin Galectin-3 Infect. Immun., August 1, 2000; 68(8): 4391 - 4398. [Abstract] [Full Text] [PDF]

				R. P. Boulianne, Y. Liu, M. Aebi, B. C. Lu, and U. Kües Fruiting body development in Coprinus cinereus: regulated expression of two galectins secreted by a non-classical pathway Microbiology, August 1, 2000; 146(8): 1841 - 1853. [Abstract] [Full Text]

				P. W. Denny, S. Gokool, D. G. Russell, M. C. Field, and D. F. Smith Acylation-dependent Protein Export in Leishmania J. Biol. Chem., April 6, 2000; 275(15): 11017 - 11025. [Abstract] [Full Text] [PDF]

				H. C. Gong, Y. Honjo, P. Nangia-Makker, V. Hogan, N. Mazurak, R. S. Bresalier, and A. Raz The NH2 Terminus of Galectin-3 Governs Cellular Compartmentalization and Functions in Cancer Cells Cancer Res., December 1, 1999; 59(24): 6239 - 6245. [Abstract] [Full Text] [PDF]

				H. Garoff, R. Hewson, and D.-J. E. Opstelten Virus Maturation by Budding Microbiol. Mol. Biol. Rev., December 1, 1998; 62(4): 1171 - 1190. [Abstract] [Full Text] [PDF]

				J. L. Dunphy, A. Balic, G. J. Barcham, A. J. Horvath, A. D. Nash, and E. N. T. Meeusen Isolation and Characterization of a Novel Inducible Mammalian Galectin J. Biol. Chem., October 6, 2000; 275(41): 32106 - 32113. [Abstract] [Full Text] [PDF]