title: Arabidopsis F-box protein containing a Nictaba-related lectin domain interacts with N-acetyllactosamine structures.
abstract: The Arabidopsis thaliana genome contains a small group of bipartite F-box proteins, consisting of an N-terminal F-box domain and a C-terminal domain sharing sequence similarity with Nictaba, the jasmonate-induced glycan-binding protein (lectin) from tobacco. Based on the high sequence similarity between the C-terminal domain of these proteins and Nictaba, the hypothesis was put forward that the so-called F-box-Nictaba proteins possess carbohydrate-binding activity and accordingly can be considered functional homologs of the mammalian sugar-binding F-box or Fbs proteins which are involved in proteasomal degradation of glycoproteins. To obtain experimental evidence for the carbohydrate-binding activity and specificity of the A. thaliana F-box-Nictaba proteins, both the complete F-box-Nictaba sequence of one selected Arabidopsis F-box protein (in casu At2g02360) as well as the Nictaba-like domain only were expressed in Pichia pastoris and analyzed by affinity chromatography, agglutination assays and glycan micro-array binding assays. These results demonstrated that the C-terminal Nictaba-like domain provides the F-box-protein with a carbohydrate-binding activity that is specifically directed against N- and O-glycans containing N-acetyllactosamine (Galβ1-3GlcNAc and Galβ1-4GlcNAc) and poly-N-acetyllactosamine ([Galβ1-4GlcNAc]n) as well as Lewis A (Galβ1-3(Fucα1-4)GlcNAc), Lewis X (Galβ1-4(Fucα1-3)GlcNAc, Lewis Y (Fucα1-2Galβ1-4(Fucα1-3)GlcNAc) and blood type B (Galα1-3(Fucα1-2)Galβ1-3GlcNAc) motifs. Based on these findings one can reasonably conclude that at least the A. thaliana F-box-Nictaba protein encoded by At2g02360 can act as a carbohydrate-binding protein. The results from the glycan array assays revealed differences in sugar-binding specificity between the F-box protein and Nictaba, indicating that the same carbohydrate-binding motif can accommodate unrelated oligosaccharides.
authors: Lab Biochemistry and Glycobiology, Dept. Molecular Biotechnology, Ghent University, Coupure links 653, 9000 Ghent, Belgium.
authors: Karolina
authors: K
authors: Stefanowicz
authors: Nausicaä
authors: N
authors: Lannoo
authors: Paul
authors: P
authors: Proost
authors: Els J M
authors: EJ
authors: Van Damme
keywords: Carbohydrate-binding; Fbs, sugar-binding F-box protein; Glycan array; LacNAc, N-acetyllactosamine; Lewis structure; Nictaba, Nicotiana tabacum agglutinin; Plant lectin; Recombinant protein expression
references: Mol Microbiol. 2000 Jun;36(6):1187-96
references: 10931272
references: Proc Natl Acad Sci U S A. 2002 Aug 20;99(17):11519-24
references: 12169662
references: Glycobiology. 2002 May;12(5):299-306
references: 12070072
references: Proc Natl Acad Sci U S A. 2004 Dec 7;101(49):17033-8
references: 15563589
references: FASEB J. 2002 Jun;16(8):905-7
references: 12039875
references: Plant Cell. 2007 Jul;19(7):2278-92
references: 17630273
references: FEBS Lett. 1997 Sep 29;415(2):186-91
references: 9350993
references: Plant Cell Physiol. 2004 Jan;45(1):83-91
references: 14749489
references: Biochem Soc Trans. 2008 Oct;36(Pt 5):843-7
references: 18793148
references: Trends Plant Sci. 2004 Oct;9(10):484-9
references: 15465683
references: Crit Rev Microbiol. 1996;22(3):139-80
references: 8894399
references: Biochem Biophys Res Commun. 2011 Oct 14;414(1):101-5
references: 21945438
references: Biochem Biophys Res Commun. 2003 Apr 25;304(1):91-7
references: 12705889
references: Nature. 1970 Aug 15;227(5259):680-5
references: 5432063
references: Curr Opin Plant Biol. 2006 Dec;9(6):631-8
references: 17005440
references: Biochem Biophys Res Commun. 2009 Mar 6;380(2):260-5
references: 19167365
references: Biochim Biophys Acta. 2010 Feb;1800(2):172-80
references: 19665047
references: Arch Virol. 2004 Jan;149(1):75-91
references: 14689277
references: J Biol Chem. 2008 May 9;283(19):12717-29
references: 18203720
references: FEBS J. 2011 Jun;278(12):2064-79
references: 21481190
references: J Biotechnol. 2010 Mar;146(1-2):54-65
references: 20083147
references: EMBO Rep. 2005 Mar;6(3):239-44
references: 15723043
references: Plant Physiol. 2003 Jan;131(1):114-28
references: 12529520
references: Plant Physiol. 1999 Oct;121(2):333-44
references: 10517824
references: Proc Natl Acad Sci U S A. 2001 Feb 27;98(5):2899-904
references: 11226338
references: Cell. 1996 Jul 26;86(2):263-74
references: 8706131
references: Biochim Biophys Acta. 1998 Mar 3;1383(1):9-36
references: 9546043
references: Plant J. 1997 Dec;12(6):1411-7
references: 9450345
references: Glycobiology. 2001 Apr;11(4):261-74
references: 11358875
references: Carbohydr Res. 2009 Aug 17;344(12):1487-93
references: 19515362
references: Planta. 1978 Jan;142(1):97-101
references: 24408004
references: Eur J Biochem. 1983 Aug 15;134(3):561-9
references: 6884347
references: Glycoconj J. 2010 Aug;27(6):613-23
references: 20721621
references: Nat Rev Mol Cell Biol. 2005 Jan;6(1):9-20
references: 15688063
references: Plant Physiol. 1999 Feb;119(2):725-34
references: 9952469
references: FEBS Lett. 2006 Nov 27;580(27):6329-37
references: 17084390
references: Plant Physiol. 2010 Jul;153(3):1345-61
references: 20442276