Expression of recombinant human GM2-activator protein in insect cells: Purification and characterization by mass spectrometry

Protein Expression and Purification

Volumn 27, Issue 2, 2003, Pages 259-266

  Wendeler, Michaela ^a   Lemm, Thorsten ^a,b   Weisgerber, Judith ^a   Hoernschemeyer, Joerg ^a   Bartelsen, Oliver ^a   Schepers, Ute ^a   Sandhoff, Konrad ^a  

^a UNIVERSITY OF BONN   (Germany)

^b ROCHE INNOVATION CENTER MUNICH   (Germany)

Author keywords

[No Author keywords available]

Indexed keywords

ANIMALS; BACULOVIRIDAE; CATIONS; CELL LINE; CELL LINE, TRANSFORMED; CHROMATOGRAPHY; CHROMATOGRAPHY, ION EXCHANGE; DNA, COMPLEMENTARY; G(M2) ACTIVATOR PROTEIN; GENETIC VECTORS; GLYCOSYLATION; HUMANS; IMMUNOBLOTTING; INSECTS; MASS SPECTROMETRY; MODELS, GENETIC; MUTATION; OLIGOSACCHARIDES; PROTEINS; RECOMBINANT PROTEINS; TRANSFECTION;

INSECTA; UNIDENTIFIED BACULOVIRUS;

EID: 0037296180     PISSN: 10465928     EISSN: None     Source Type: Journal    
DOI: 10.1016/S1046-5928(02)00599-5     Document Type: Article

References (21)

1. W. Fürst, K. Sandhoff, Activator proteins and topology of lysosomal sphingolipid catabolism, Biochim. Biophys. Acta 1126 (1992) 1-6.
2. T. Kolter, K. Sandhoff, Sphingolipids - Their metabolic pathways and the pathobiochemistry of neurodegenerative diseases, Angew. Chem. Int. Ed. Engl. 38 (1999) 1532-1568.
3. T. Nakano, K. Sandhoff, J. Stümper, H. Christomanou, K. Suzuki, Structure of full-length cDNA coding for sulfatide activator, a co-β-glucosidase and two other homologous proteins: two alternate forms of the sulfatide activator, J. Biochem. (Tokyo) 105 (1989) 152-154.
4. E.M. Meier, G. Schwarzmann, W. Fürst, K. Sandhoff, The human GM2 activator protein: a substrate specific cofactor of hexosaminidase A, J. Biol. Chem. 266 (1991) 1879-1887.
5. S.C. Li, S. Sonnino, G. Tettamanti, Y.T. Li, Characterization of a nonspecific activator protein for the enzymatic hydrolysis of glycolipids, J. Biol. Chem. 263 (1988) 6588-6591.
6. H.J. Kytzia, K. Sandhoff, Evidence for two different active sites on human hexosaminidase - interaction of GM2 activator protein with hexosaminidase A, J. Biol. Chem. 260 (1985) 7568-7572.
7. T. Kolter, K. Sandhoff, Enzymology of lysosomal glycolipid catabolism, Trends Glycosci. Glycotechnol. 10 (1998) 455-468.
8. A. Giehl, T. Lemm, O. Bartelsen, K. Sandhoff, A. Blume, Interaction of the GM2 activator protein with phospholipidganglioside bilayer membranes and with monolayers at the airwater interface, Eur. J. Biochem. 261 (1999) 650-658.
9. G. Wilkening, T. Linke, K. Sandhoff, Lysosomal degradation on vesicular membrane surfaces, J. Biol. Chem. 273 (1998) 30271-30278.
10. G.J. Glombitza, E. Becker, H.W. Kaiser, K. Sandhoff, Biosynthesis, processing, and intracellular transport of GM2 activator protein in human epidermal keratinocytes, J. Biol. Chem. 272 (1997) 5199-5207.
11. E. Conzelmann, K. Sandhoff, AB variant of infantile GM2 gangliosidosis: deficiency of a factor necessary for stimulation of hexosaminidase A-catalyzed degradation of ganglioside GM2 and glycolipid GA2, Proc. Natl. Acad. Sci. USA 75 (1978) 3979-3983.
12. H. Klima, A. Klein, G. van Echten, G. Schwarzmann, K. Suzuki, K. Sandhoff, Over-expression of a functionally active human GM2-activator protein in Escherichia coli, Biochem. J. 292 (1993) 571-576.
13. Y.Y. Wu, J.M. Lockyer, E. Sugiyama, N.V. Pavlova, Y.T. Li, S.C. Li, Expression and specificity of human GM2 activator protein, J. Biol. Chem. 269 (1994) 16276-16283.
14. C.S. Wright, S.C. Li, F. Rastinejad, Crystal structure of human GM2-activator protein with a novel β-cup topology, J. Mol. Biol. 304 (2000) 411-422.
15. J. Sambrook, E.F. Fritsch, T.L. Maniatis, Molecular Cloning: A Laboratory Manual, second ed., Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY, 1989.
16. N.H. Packer, M.A. Lawson, D.R. Jardine, J.W. Redmond, A general approach to desalting oligosaccharides released from glycoproteins, Glycoconj. J. 15 (1998) 737-747.
17. E. Conzehnann, K. Sandhoff, Purification and characterization of an activator protein for the degradation of glycolipids GM2 and GA2 by hexosaminidase A, Hoppe-Seyler's Z. Physiol. Chem. 360 (1979) 1837-1849.
18. D.R. O'Reilly, L.K. Miller, V.A. Luckow, Baculovirus Expression Vectors: A Laboratory Manual, W.H. Freeman, New York, 1992.
19. D.L. Jarvis, Baculovirus expression vectors, in: L.K. Miller (Ed.), The Baculoviruses, Plenum Press, New York, 1997, pp. 389-431.
20. D.L. Jarvis, C. Weinkauf, L.A. Guarino, Immediate-early baculovirus vectors for foreign gene expression in transformed or infected insect cells, Protein Expr. Purif. 8 (1996) 191-203.
21. D.D. Hegedus, T.A. Pfeifer, J. Hendry, D.A. Theilmann, T.A. Grigliatti, A series of broad host range shuttle vectors for constitutive and inducible expression of heterologous proteins in insect cell lines, Gene 207 (1998) 241-249.