Abstract
Endo-β-galactosidases cleave the internal β-linked galactosyl residues of sugar chains in glycoconjugates and release oligosaccharides as their products. Based on the oligosaccharide products, the microbial endo-β-galactosidases [1] can be divided into four classes, of which the products are, respectively: (a) GlcNAcβ1 → 3Gal, (b) Galα1 → 3Gal, (c) blood group A-trisaccharide and B-trisaccharide, and (d) GlcNAcα1 → 4Gal. Endo-β-galactosidases, as well as other glycosidases, are often present as contaminants in commercial enzyme preparations. Their presence as contaminants is not easily recognized or clearly indicated. The use of an enzyme preparation with unknown contaminants can give unexplainable results or cause misinterpretation. However, the enzyme contaminants may sometimes provide opportunities for new discoveries. Endo-ABase was discovered through the use of an impure commercial clostridial sialidase preparation.
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Abbreviations
- A+-PGM:
-
Blood group A+ porcine gastric mucin
- B+-HOCG:
-
Blood group B+ human ovarian cyst glycoprotein
- A-Tri:
-
GalNAcα1 → 3(Fucα1 → 2)Gal
- A-Tetra:
-
GalNAcα1 → 3(Fucα1 → 2) Galβ1 → 4Glc
- A-Penta:
-
GalNAcα1 → 3(Fucα1 → 2)Galβ1 → 4(Fucα1 → 3)Glc
- A-Hexa:
-
GalNAcα1 → 3-(Fucα1 → 2)Galβ1 → 3GlcNAcβ1 → 3Galβ1 → 4Glc
- B-Tri:
-
Galα1 → 3(Fucα1 → 2)Gal
- B-Penta:
-
Galα1 → 3-(Fucα1 → 2)Galβ1 → 4(Fucα1 → 3)Glc
- Endo-ABase:
-
Blood group A and B cleaving endo-β-galactosidase
- nEndo-ABase and rEndo-ABase:
-
The native and the recombinant Endo-ABase respectively
- FPLC:
-
Fast protein liquid chromatography
- TLC:
-
Thin-layer chromatography
- ConA:
-
Concanavalin A
- LB:
-
Luria–Bertani medium
- CNBr:
-
Cyanogen bromide
- DEAE:
-
Diethylaminoethyl
- SP:
-
Sulfopropyl
- Ni-NTA:
-
Ni-nitrilotri-acetic acid
- CV:
-
Column volume
- SSC:
-
17.5% Sodium chloride/8.8% sodium citrate, pH 7.0
- SM:
-
0.58% Sodium chloride/0.20% magnesium sulfate/50 mM Tris–HCl, pH 7.5/0.01% gelatin
- RBC:
-
Human red blood cells
- FRP:
-
Fucolectin-related protein
- FITC:
-
Fluorescene-5-isothiocyanate
- FACS:
-
Fluorescene-activated cell sorter
References
Ashida H, Li SC, Li YT (2006) An unusual GlcNAcα1-4Gal releasing endo-β-galactosidase. In: Endo M, Hasse S, Yamamoto K, Takagaki K (eds) Endoglycosidases, biochemistry, biotechnology, application. Kodansha/Springer, Tokyo, Japan
Chou MY, Li SC, Kiso M, Hasegawa A, Li YT (1994) Purification and characterization of sialidase L, NeuAcα2 → 3Gal-specific sialidase. J Biol Chem 269:18821–18826
Cassidy JT, Jourdian GW, Roseman S (1965) The sialic acids IV. Purification and properties of sialidase from Clostridium perfringens. J Biol Chem 240:3501–3506
Corfield T (1992) Bacterial sialidases – roles in pathogenicity and nutrition. Glycobiology 2:509–521
Hatton MWC, Regoeczi E (1973) A simple method for the purification of commercial neuraminidase preparations free from proteases. Biochim Biophys Acta 327:114–120
Chien SF, Yevich SJ, Li SC, Li YT (1975) Presence of endo-β-N-acetylglucosaminidase and protease activities in the commercial neuraminidase preparations isolated from Clostridium perfringens. Biochem Biophys Res Commun 65:683–691
Boyle MDP, Ohanian SH, Borsos T (1976) Lysis of tumor cells by antibody and complement VI. Enhanced killing of enzyme pretreated tumor cells. J Immunol 116:661–668
Kraemer PM (1968) Cytotoxic, hemolytic and phospholipase contaminants of commercial neuraminidases. Biochim Biophys Acta 167:205–208
Den H, Malinzak DA, Rosenberg A (1975) Cytotoxic contaminants in commercial Clostridium perfringens neuraminidase preparations purified by affinity chromatography. J Chromatogr 111:217–222
Trams EG, Lauter CJ, Banfield WG (1976) On the activation of plasma membrane ecto-enzymes by treatment with neuraminidase. J Neurochem 27:1035–1042
Ashida H, Anderson KM, Nakayama J, Maskos K, Chou CW, Cole R, Li SC, Li YT (2001) A novel endo-β-galactosidase from Clostridium perfringens that liberates the disacchride GlcNAcα1 → 4Gal from glycans specifically expressed in the gastric gland mucous cell-type mucin. J Biol Chem 276:28226–28232
Anderson KM, Ashida H, Maskos K, Dell A, Li SC, Li YT (2005) A Clostridial endo-β-galactosidase that cleaves both blood group A and B glycotopes. J Biol Chem 280:7720–7728
Anderson KM, Li SC, Li YT (2000) Diphenylamine-aniline-phosphoric acid reagent, a versatile spray reagent for revealing glycoconjugates on thin layer chromatography plates. Anal Biochem 287:337–339
Cassidy JT, Jourdian GW, Roseman S (1966) Sialidase from Clostridium perfringens. Methods Enzymol 8:680–685
Charbonneau H (1989) CNBr digest. In: Matsudaira PT (ed) A practical guide to protein and peptide purification for microsequencing. Academic, San Diego, CA
Ausubel FM, Brent R, Kingston RE, Moore DD, Seidman JG, Smith JA, Struhl K (1991) Current protocols in molecular biology. Wiley, New York
Ashida H, Maskos K, Li SC, Li YT (2002) Characterization of a novol endo-β-galactosidase specific for releasing the disaccharide GlcNAcα1 → 4Gal from glycoconjugates. Biochemistry 41:2388–2395
Hoffmann A, Roeder RG (1991) Purification of His-tagged proteins in non-denaturing conditions suggests a convenient method for protein fraction studies. Nucleic Acids Res 19:6337–6338
Clausen H, Hakomori SI (1989) ABH and related histo-blood group antigens; immunochemical differences in carrier isotypes and their distribution. Vox Sang 56:1–20
Hoskins LC, Larson G, Naff GB (1995) Blood group A immunodeterminants on human red cells differ in biologic activity and sensitivity to α-N-acetylgalactosaminidase. Transfusion 35:813–821
Kobayashi T, Liu D, Ogawa H, Miwa Y, Nagasaka T, Maruyama S, Li YT, Onishi A, Kuzuya T, Kadomatsu K, Uchida K, Nakao A (2007) Alternative strategy for overcoming ABO-incompatibility. Transplantation 83:1284–1286
Shimizu T, Ohtani K, Hirakawa H, Ohshima K, Yamashita A, Shiba T, Ogasawara N, Hattori M, Kuhara S, Hayashi H (2002) Complete genome sequence of Clostridium perfringens, an anaerobic flesh-eater. Proc Natl Acad Sci USA 99:996–1001
Tettelin H et al (2001) Complete genome sequence of a virulent isolate of Streptococcus pneumoniae. Science 293:498–506
Takasaki S, Kobata A (1976) Purification and characterization of an endo-β-galactosidase produced by Diplococcus pneumoniae. J Biol Chem 251:3603–3609
Leng L, Zhu A, Zhang Z, Hurst R, Goldstein J (1998) Cloning, functional expression and purification of endo-β-galactosidase from Flavobacterium keratolyticus. Gene 222:187–194
Ogawa H, Muramatsu H, Kobayashi T, Morozumi K, Yokoyama I, Kurosawa N, Nakao A, Muramatsu T (2000) Molecular cloning of endo-β-galactosidase C and its application in removing α-galactosyl xenoantigen from blood vessels in the pig kidney. J Biol Chem 275:19368–19374
Greenwell P (1997) Blood group antigens: molecules seeking a function? Glycoconj J 14:159–173
Acknowledgments
We thank the late Professor Winifred M. Watkins for the generous gift of the HOCG samples used in this work. Thanks are also due to Dr. Bernard Henrissat for his invaluable advice on the classification of GHs. This work was supported by National Institutes of Health grant NS 09626.
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Li, SC., Anderson, K.M., Li, YT. (2011). A Unique Endo-β-Galactosidase that Cleaves Both Blood Group A and B Glycotopes. In: Wu, A. (eds) The Molecular Immunology of Complex Carbohydrates-3. Advances in Experimental Medicine and Biology, vol 705. Springer, Boston, MA. https://doi.org/10.1007/978-1-4419-7877-6_5
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DOI: https://doi.org/10.1007/978-1-4419-7877-6_5
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