Caracterización enzimática de la α-1,3- galactosiltransferasa bovina. Validación de un ensayo radiométrico y mecanismo cinético

Ana Monegal; Vincent Bulone; Antoni Planas

Caracterización enzimática de la α-1,3- galactosiltransferasa bovina. Validación de un ensayo radiométrico y mecanismo cinético

Translated title of the contribution: Enzymatic characterization of bovine α-1,3-galactosyltransferase. Validation of a radiometric assay and kinetic mechanism

Ana Monegal, Vincent Bulone, Antoni Planas

IQS School of Engineering

Research output: Indexed journal article › Article › peer-review

1 Citation (Scopus)

Abstract

α3-Galactosyltransferase (α3GT) transfers galactose from UDP-Gal (sugar nucleotide donor) to the N-acetyllactosaminyl or lactosyl terminal groups of glycoproteins and glycolipids, catalyzing the formation of an α-1,3 glycosidic bond. The terminal saccharide Galα3NAcGalβ 4Glu-R is the main antigenic determinant responsible of the hyperaccute rejection in xenotransplantation. A radiometric assay for the determination of α3GT activity is implemented and validated. The recombinant enzyme (catalytic domain) expressed in Eschericia coli follows a bi bi sequential ordered kinetic mechanism with binding of donor substrate (UDP-Gal) first and acceptor substrate to form a productive ternary complex. K _M values are 30 μM for UDP-Gal, and 1.2 mM for lactose.

Translated title of the contribution	Enzymatic characterization of bovine α-1,3-galactosyltransferase. Validation of a radiometric assay and kinetic mechanism
Original language	Spanish
Pages (from-to)	505-512
Number of pages	8
Journal	Afinidad
Volume	62
Issue number	519
Publication status	Published - Sept 2005

Cite this

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title = "Caracterizaci{\'o}n enzim{\'a}tica de la α-1,3- galactosiltransferasa bovina. Validaci{\'o}n de un ensayo radiom{\'e}trico y mecanismo cin{\'e}tico",

abstract = "α3-Galactosyltransferase (α3GT) transfers galactose from UDP-Gal (sugar nucleotide donor) to the N-acetyllactosaminyl or lactosyl terminal groups of glycoproteins and glycolipids, catalyzing the formation of an α-1,3 glycosidic bond. The terminal saccharide Galα3NAcGalβ 4Glu-R is the main antigenic determinant responsible of the hyperaccute rejection in xenotransplantation. A radiometric assay for the determination of α3GT activity is implemented and validated. The recombinant enzyme (catalytic domain) expressed in Eschericia coli follows a bi bi sequential ordered kinetic mechanism with binding of donor substrate (UDP-Gal) first and acceptor substrate to form a productive ternary complex. K M values are 30 μM for UDP-Gal, and 1.2 mM for lactose.",

keywords = "Activity, Galactosyltransferase, Kinetic mechanism, Radiometric assay, Sequential ordered mechanim, Validation",

author = "Ana Monegal and Vincent Bulone and Antoni Planas",

year = "2005",

month = sep,

language = "Espa{\~n}ol",

volume = "62",

pages = "505--512",

journal = "Afinidad",

issn = "0001-9704",

publisher = "Asociacion de Quimicos del Instituto Quimico de Sarria",

number = "519",

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TY - JOUR

T1 - Caracterización enzimática de la α-1,3- galactosiltransferasa bovina. Validación de un ensayo radiométrico y mecanismo cinético

AU - Monegal, Ana

AU - Bulone, Vincent

AU - Planas, Antoni

PY - 2005/9

Y1 - 2005/9

N2 - α3-Galactosyltransferase (α3GT) transfers galactose from UDP-Gal (sugar nucleotide donor) to the N-acetyllactosaminyl or lactosyl terminal groups of glycoproteins and glycolipids, catalyzing the formation of an α-1,3 glycosidic bond. The terminal saccharide Galα3NAcGalβ 4Glu-R is the main antigenic determinant responsible of the hyperaccute rejection in xenotransplantation. A radiometric assay for the determination of α3GT activity is implemented and validated. The recombinant enzyme (catalytic domain) expressed in Eschericia coli follows a bi bi sequential ordered kinetic mechanism with binding of donor substrate (UDP-Gal) first and acceptor substrate to form a productive ternary complex. K M values are 30 μM for UDP-Gal, and 1.2 mM for lactose.

AB - α3-Galactosyltransferase (α3GT) transfers galactose from UDP-Gal (sugar nucleotide donor) to the N-acetyllactosaminyl or lactosyl terminal groups of glycoproteins and glycolipids, catalyzing the formation of an α-1,3 glycosidic bond. The terminal saccharide Galα3NAcGalβ 4Glu-R is the main antigenic determinant responsible of the hyperaccute rejection in xenotransplantation. A radiometric assay for the determination of α3GT activity is implemented and validated. The recombinant enzyme (catalytic domain) expressed in Eschericia coli follows a bi bi sequential ordered kinetic mechanism with binding of donor substrate (UDP-Gal) first and acceptor substrate to form a productive ternary complex. K M values are 30 μM for UDP-Gal, and 1.2 mM for lactose.

KW - Activity

KW - Galactosyltransferase

KW - Kinetic mechanism

KW - Radiometric assay

KW - Sequential ordered mechanim

KW - Validation

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M3 - Artículo

AN - SCOPUS:33745042465

SN - 0001-9704

VL - 62

SP - 505

EP - 512

JO - Afinidad

JF - Afinidad

IS - 519

ER -

Caracterización enzimática de la α-1,3- galactosiltransferasa bovina. Validación de un ensayo radiométrico y mecanismo cinético

Abstract

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