Abstract
The purification of fucose-binding lectins from the liver of striped bass (Morone saxatilis), a teleost fish, and the identification of a novel lectin sequence motif led to the identification of a new family of lectins, the F-type lectins (FTLs) (see overview of the FTL family in Chapter 23). Isolation and purification of these proteins from liver extracts of striped bass was accomplished by affinity chromatography and size exclusion, and their identification as FTLs, by direct Edman sequencing, and protein, transcript, and gene sequence analysis. The development of specific antibodies against the M. saxatilis FTL provided an additional tool for the identification of FTLs. These methods have been successfully used for the purification of the FTL family members from tissues and body fluids of various animal species. Production and characterization of FTLs has been facilitated by the expression of the recombinant proteins. In this chapter, the biochemical characterization of FTLs is focused on the analysis of their carbohydrate specificity.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
Similar content being viewed by others
Abbreviations
- BSA:
-
Bovine serum albumin
- BSM:
-
Bovine submaxillary mucin
- CBB:
-
Coomassie Brilliant Blue
- ELISA:
-
Enzyme-linked immunosorbent assay
- FBD:
-
Fucose-binding domains
- FTL:
-
F-type lectin
- Igs:
-
Immunoglobulins
- IPTG:
-
Isopropyl β-D-thiogalactoside
- MsaFBP32:
-
M. saxatilis fucose-binding protein 32 kDa
- NCFG:
-
National Center for Functional Glycomics
- OD:
-
Optical density
- OSM:
-
Ovine submaxillary mucin
- PBS:
-
Phosphate-buffered saline
- PMSF:
-
Phenylmethylsulfonyl fluoride
- PSM:
-
Porcine stomach mucin
- RBC:
-
Red blood cells
- RP-HPLC:
-
Reverse phase-HPLC
- SEC:
-
Size exclusion chromatography
- TFA:
-
Trifluoroacetic acid
References
Vasta GR, Ahmed H (2008) Animal Lectins: a functional view. Taylor & Francis, CRC Press, Boca Raton, p 558
Vasta GR, Hunt JC, Marchalonis JJ et al (1986) Galactosyl-binding lectins from the tunicate Didemnum candidum. Purification and physicochemical characterization. J Biol Chem 261:9174–9181
Vasta GR, Marchalonis JJ (1986) Galactosyl-binding lectins from the tunicate Didemnum candidum. Carbohydrate specificity and characterization of the combining site. J Biol Chem 261:9182–9186
Ahmed H, Fink NE, Pohl J et al (1996) Galectin-1 from bovine spleen: biochemical characterization, carbohydrate specificity and tissue-specific isoform profiles. J Biol Chem 120:1007–1019
Ahmed H, Du SJ, O’leary N et al (2004) Biochemical and molecular characterization of galectins from zebrafish (Danio rerio): notochord-specific expression of a prototype galectin during early embryogenesis. Glycobiology 14:219–232
Bianchet MA, Odom EW, Vasta GR et al (2002) A novel fucose recognition fold involved in innate immunity. Nat Struct Biol 9:628–634
Odom EW, Vasta GR (2006) Characterization of a binary tandem domain F-type lectin from striped bass (Morone saxatilis). J Biol Chem 281:1698–1713
Tasumi S, Vasta GR (2007) A galectin of unique domain organization from hemocytes of the eastern oyster (Crassostrea virginica) is a receptor for the protistan parasite Perkinsus marinus. J Immunol 179:3086–3098
Latha VL, Rao RN, Nadimpalli SK (2006) Affinity purification, physicochemical and immunological characterization of a galactose-specific lectin from the seeds of Dolichos lablab (Indian lablab beans). Protein Expr Purif 45:296–306
Mo H, Meah Y, Moore JG et al (1999) Purification and characterization of Dolichos lablab lectin. Glycobiology 9:173–179
Odom EW (2004). F-type Lectins: biochemical, genetic and structural characterization of a novel Lectin family in lower vertebrates. Ph.D. thesis. Baltimore, MD: University of Maryland, MEES Program
Vasta GR, Ahmed H, Bianchet MA et al (2012) Diversity in recognition of glycans by F-type lectins and galectins: molecular, structural, and biophysical aspects. Ann N Y Acad Sci 1253:E14–E26
Vasta GR, Amzel LM, Bianchet MA et al (2017) F-type Lectins: a highly diversified family of Fucose-binding proteins with a unique sequence motif and structural fold, involved in self/non-self-recognition. Front Immunol 8:1648
Tennent GA, Pepys MB (1994) Glycobiology of the pentraxins. Biochem Soc Trans 22:74–79
Hermanson GT, Mallia AK, Smith PK (1992) Immobilized affinity ligand techniques. Academic Press, San Diego
Feng C, Ghosh A, Amin MN et al (2015) Galectin CvGal2 from the eastern oyster (Crassostrea virginica) displays unique specificity for ABH blood group oligosaccharides and differentially recognizes sympatric Perkinsus species. Biochemistry 54:4711–4730
Feng C, Ghosh A, Amin MN et al (2013) The galectin CvGal1 from the eastern oyster (Crassostrea virginica) binds to blood group a oligosaccharides on the hemocyte surface. J Biol Chem 288:24394–24409
Chevallet M, Luche S, Rabilloud T (2006) Silver staining of proteins in polyacrylamide gels. Nat Protoc 1:1852–1858
Hewick RM, Hunkapiller MW, Hood LE et al (1981) A gas-liquid solid phase peptide and protein sequenator. J Biol Chem 256:7990–7997
Pohl J (1994) Sequence analysis of peptide resins from Boc/benzyl solid-phase synthesis. Methods Mol Biol 36:107–129
Coligan JE (1995) Current protocols in protein science. Wiley, Brooklyn, NY
Nielsen H, Engelbrecht J, Brunak S et al (1997) Identification of prokaryotic and eukaryotic signal peptides and prediction of their cleavage sites. Protein Eng 10:1–6
Watkins WM, Morgan WT (1952) Neutralization of the anti-H agglutinin in eel serum by simple sugars. Nature 169:825–826
Acknowledgments
The author’s research reviewed herein was supported by Grants IOS 1050518, IOB-0618409, MCB 0077928, and IOS-0822257 from the National Science Foundation, and Grant R01GM070589 from the National Institutes of Health to G.R.V.
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2020 Springer Science+Business Media, LLC, part of Springer Nature
About this protocol
Cite this protocol
Feng, C., Vasta, G.R. (2020). Purification and Biochemical Characterization of Selected F-Type Lectins. In: Hirabayashi, J. (eds) Lectin Purification and Analysis. Methods in Molecular Biology, vol 2132. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-0430-4_24
Download citation
DOI: https://doi.org/10.1007/978-1-0716-0430-4_24
Published:
Publisher Name: Humana, New York, NY
Print ISBN: 978-1-0716-0429-8
Online ISBN: 978-1-0716-0430-4
eBook Packages: Springer Protocols