α1-6 fucosyltransferase

Chemistry: molecular biology and microbiology – Enzyme – proenzyme; compositions thereof; process for... – Transferase other than ribonuclease

Reexamination Certificate

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C435S004000, C435S006120, C435S069100, C435S183000, C435S252300, C435S320100, C536S023200, C536S023400, C536S023500, C536S023700

Reexamination Certificate

active

10844432

ABSTRACT:
An isolated, porcine α1-6 fucosyltransferase having the following physico-chemical properties:(1) action: transferring fucose from guanosine diphosphate-fucose to a hydroxy group at 6-position of GluNAc closest to R of a receptor (GlcNAcβ1-2Manα1-6)(GlcNAcβ1-2Manα1-3)Manβ1-4GlcNAcβ1-4GlucNAc-R wherein R is an asparagine residue or a peptide chain carrying said residue, whereby to form (GlcNAcβ1-2Manα1-6)-(GlcNAcβ1-2Manα1-3)Manα1-4GlcNAcβ1-4(Fucα1-6)GlucNAc-R(2) optimum pH: about 7.0(3) pH stability: retains activity after 5 hours of treatment at 4° C. at a pH range of 4.0-10.0(4) optimum temperature: about 30-37° C.(5) inhibition or activation: no requirement for divalent metal for expression of activity; no inhibition of activity in the presence of 5 mM EDTA(6) molecular weight: about 60,000 by SDS-polyacrylamide gel electrophoresisis provided. An isolated polynucleotide encoding porcine α1-6 fucosyltransferase is also provided. Expression vectors comprising a polynucleotide encoding porcine α1-6 fucosyltransferase are also provided.

REFERENCES:
Longmore et al., Carbohydrate Res., 1982, vol. 100 :365-392.
Uozumi N. et al.,Purification and cDNA cloning of porcine brain, 1996, J. Biol. Chem, 271(44) 27810-27817.
Uozumi N. et al.,A fluorescent assay method for GDP-L-Fuc: N-acetyl-beta-D-glucosaminide alpha-1-6fucosyltransferease activity, involving high performance liquid chromatorgraphy, 1996, J. Biochem., 120(2) 385-392.
Voynow J. A. et al.,Purification and characterization of GDP-L-fucose-N-acetyl-beta-D-glucosaminide alpha-1-6 fucosyltransferase from cultured human skin fibroblasts requirement of a specific biantennary oligosaccharide as substrate, 1991, J. Biol. Chem., 266(32) 21572-21577.
“Product Identification and Substrate Specificity Studies of the GDP-L Fucose 2 Acetamido-2-Deoxy-Beta-D Glucoside Fucose Asparagine Linked N Acetyl Glucosamine 6-Alpha-L Fucosyl Transferase in a Golgi-Rich Fraction from Porcine Liver” Carbohydrate Research, vol. 100, 1982 pp. 365-392 by G.D. Longmore, et al.
“The alpha1-6-fucosyltransferase gene and its biological significance” Biochimica ET Biophysica ACTA, vol. 1473, No. 1, Dec. 6, 1999, pp. 9-20 by Eiji Miyoshi et al.
Hillier et al. GenBank Accession No. AA057866, dated Sep. 18, 1996.
Uozumi, N. et al., “Purification and Characteristics of α1-6 Fucosyltransferase (α1-6FucT) Derived from Porcine Brain,”in The Abstract for the 68thMeeting of the Japanese Biochemical Society, Biochemistry67(7): abstr. No. 4053 (1995).

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