N-acetylglucosaminyl transferase gene coding therefor and proces

Details N-acetylglucosaminyl transferase gene coding therefor and proces N-acetylglucosaminyl transferase gene coding therefor and proces

1997-08-14

1998-11-10

Patterson, Jr., Charles L.

Chemistry: molecular biology and microbiology

Enzyme , proenzyme; compositions thereof; process for...

Transferase other than ribonuclease

4353201, 4352523, 536 232, C12N 918, C12N 1554

Patent

active

058342842

ABSTRACT:
A .beta.1,6-N-acetylglucosaminyl transferase having the following properties:
(1) Action: it transfers N-acetylglucosamine from UDP-N-acetylglucosamine to .alpha.-6-D-mannoside;
(2) Substrate specificity: it shows a reactivity of about 79% for GnGnF-bi-PA, about 125% for GnGnGn-tri-PA and about 66% for GnM-Pa, when taking a reactivity for GnGn-bi-PA as 100%;
(3) Optimum pH: 6.2 to 6.3;
(4) Inhibition, Activation and Stability: Mn.sup.2+ is not necessary for expression of activity, and the activity is not inhibited in the presence of 20 mM EDTA;
(5) Molecular weight: about 73,000 as determined by SDS-PAGE in the absence of reducing agent; and about 73,000 and about 60,000 as determined in the presence of a reducing agent;
(6) Km value: 133 .mu.M and 3.5 mM for acceptor GnGn-bi-PA and donor UDP-GlcNAc, respectively; and
(7) It includes the following peptide fragments:

REFERENCES:
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Mohamed G. Shoreibach et al., "Purification and Characterization of Rat Kidney UDP-N-acetylglucosamine: .alpha.-6-D-Mannoside .beta.-1,6-N-Acetylglucosaminyltransferase", The Journal of Biological Chemistry, pp. 2920-2927, vol. 267, No. 5, Feb. 15, 1992, Baltimore, Maryland.
N. Fregien et al., "Isolation of Putative cDNAs which Encode N-Acetyglucosaminyltransferase V. Activity", The FASEB Journal, p. A1979, Abstract No. 1664, vol. 4, No. 5, Mar. 1990, Bethesda, Maryland.
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R. Cummings et al., "A Mouse Lymphoma Cell Line Resistant to the Leukoagglutinating Lectin from Phaseolus vulgaris Is Deficient in UDP-GIcNAc:.alpha.-D-mannoside .beta.1, 6 N-Acetylglucosaminyltransferase", J. Biol. Chem., vol. 257, No. 22, pp. 13421-13427, 1982.
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J.W. Dennis et al., "Oncodevelopmental Expression of -GIcNac.beta.1-6M and .alpha.1-6Man.beta.1-Branched Asparagine-linked Oligosaccharides in Murine Tissues and Human Breast Carcinomas", Cancer Res., vol. 49, pp. 945-950, Feb. 1989.
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A. Nishikawa et al., "Determination of N-Acetylgluscosaminyltransferases III, IV and V in Normal and Hepatoma Tissues of Rats", Biochimica et Biophysica Acta, vol. 1035, pp. 313-318, 1990.
N. Taniguchi et al., "Glycosyltransferase Assys Using Pyridylaminated Acceptors: N-Acetylglucosaminyltransferase III, IV, and V", Methods Enzymol., vol. 179, pp. 397-408, 1989.
M. Redinbaugh, "Adaptation of the Bicinchoninic Acid protein Assay for Use With Microtiter Plates and Sucrose Gradient Factions", Analytical Biochemistry, vol. 153, pp. 267-271, 1986.
H. Schachter et al., "Molecular Cloning Of Human and Rabbit UDP-N-acetylglucosamine: .alpha.-3-D-mannoside .beta.1,2-N-acetylglucosaminyltransferase I", Biochemical Society Transactions, 19(3), pp. 645-648. (1991).

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