Purification and characterization of CMP-NeuAc:GM1 (Galβ1-4Ga1NAc) α2-3 sialyltransferase from rat brain

Tian Jue Gu, Xin Bin Gu, Toshio Ariga, Robert K. Yu

Research output: Contribution to journalArticle

12 Citations (Scopus)

Abstract

A CMP-NeuAc:GM1 α2-3sialyltransferase (GD1a synthase, 2.4.99.2) has been purified from the Triton extract of rat brain. The enzyme was purified and resolved by affinity chromatography on CDP-Sepharose column by a linear NaCl gradient elution. Final purification was achieved by elution from a 'GM1-acid'-Sepharose column. SDS-PAGE of the enzyme revealed a single protein band with an apparent Mr 44 kDa. It catalyzed specifically the sialylation of GD1b, GM1 and asialo-GM1. Enzyme products were identified by TLC in three different solvent systems, The Km value for GM1 was 7.5 × 10-2 M, and for CMP-NeuAc it was 6.5 × 10-5 M.

Original languageEnglish (US)
Pages (from-to)83-86
Number of pages4
JournalFEBS Letters
Volume275
Issue number1-2
DOIs
StatePublished - Nov 26 1990
Externally publishedYes

Fingerprint

Sialyltransferases
Cytidine Monophosphate
Purification
Rats
Brain
Sepharose
Enzymes
Cytidine Diphosphate
Affinity chromatography
Affinity Chromatography
Polyacrylamide Gel Electrophoresis
Acids
Proteins

Keywords

  • GM1
  • Ganglioside
  • Glycolipid
  • Glycosyltransferase
  • Sialyltransferase

ASJC Scopus subject areas

  • Structural Biology
  • Biophysics
  • Biochemistry
  • Molecular Biology
  • Genetics
  • Cell Biology

Cite this

Purification and characterization of CMP-NeuAc:GM1 (Galβ1-4Ga1NAc) α2-3 sialyltransferase from rat brain. / Gu, Tian Jue; Gu, Xin Bin; Ariga, Toshio; Yu, Robert K.

In: FEBS Letters, Vol. 275, No. 1-2, 26.11.1990, p. 83-86.

Research output: Contribution to journalArticle

@article{5c242a897c074223ab89691f7e23cd8f,
title = "Purification and characterization of CMP-NeuAc:GM1 (Galβ1-4Ga1NAc) α2-3 sialyltransferase from rat brain",
abstract = "A CMP-NeuAc:GM1 α2-3sialyltransferase (GD1a synthase, 2.4.99.2) has been purified from the Triton extract of rat brain. The enzyme was purified and resolved by affinity chromatography on CDP-Sepharose column by a linear NaCl gradient elution. Final purification was achieved by elution from a 'GM1-acid'-Sepharose column. SDS-PAGE of the enzyme revealed a single protein band with an apparent Mr 44 kDa. It catalyzed specifically the sialylation of GD1b, GM1 and asialo-GM1. Enzyme products were identified by TLC in three different solvent systems, The Km value for GM1 was 7.5 × 10-2 M, and for CMP-NeuAc it was 6.5 × 10-5 M.",
keywords = "GM1, Ganglioside, Glycolipid, Glycosyltransferase, Sialyltransferase",
author = "Gu, {Tian Jue} and Gu, {Xin Bin} and Toshio Ariga and Yu, {Robert K.}",
year = "1990",
month = "11",
day = "26",
doi = "10.1016/0014-5793(90)81444-S",
language = "English (US)",
volume = "275",
pages = "83--86",
journal = "FEBS Letters",
issn = "0014-5793",
publisher = "Elsevier",
number = "1-2",

}

TY - JOUR

T1 - Purification and characterization of CMP-NeuAc:GM1 (Galβ1-4Ga1NAc) α2-3 sialyltransferase from rat brain

AU - Gu, Tian Jue

AU - Gu, Xin Bin

AU - Ariga, Toshio

AU - Yu, Robert K.

PY - 1990/11/26

Y1 - 1990/11/26

N2 - A CMP-NeuAc:GM1 α2-3sialyltransferase (GD1a synthase, 2.4.99.2) has been purified from the Triton extract of rat brain. The enzyme was purified and resolved by affinity chromatography on CDP-Sepharose column by a linear NaCl gradient elution. Final purification was achieved by elution from a 'GM1-acid'-Sepharose column. SDS-PAGE of the enzyme revealed a single protein band with an apparent Mr 44 kDa. It catalyzed specifically the sialylation of GD1b, GM1 and asialo-GM1. Enzyme products were identified by TLC in three different solvent systems, The Km value for GM1 was 7.5 × 10-2 M, and for CMP-NeuAc it was 6.5 × 10-5 M.

AB - A CMP-NeuAc:GM1 α2-3sialyltransferase (GD1a synthase, 2.4.99.2) has been purified from the Triton extract of rat brain. The enzyme was purified and resolved by affinity chromatography on CDP-Sepharose column by a linear NaCl gradient elution. Final purification was achieved by elution from a 'GM1-acid'-Sepharose column. SDS-PAGE of the enzyme revealed a single protein band with an apparent Mr 44 kDa. It catalyzed specifically the sialylation of GD1b, GM1 and asialo-GM1. Enzyme products were identified by TLC in three different solvent systems, The Km value for GM1 was 7.5 × 10-2 M, and for CMP-NeuAc it was 6.5 × 10-5 M.

KW - GM1

KW - Ganglioside

KW - Glycolipid

KW - Glycosyltransferase

KW - Sialyltransferase

UR - http://www.scopus.com/inward/record.url?scp=0025153401&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0025153401&partnerID=8YFLogxK

U2 - 10.1016/0014-5793(90)81444-S

DO - 10.1016/0014-5793(90)81444-S

M3 - Article

C2 - 2262006

AN - SCOPUS:0025153401

VL - 275

SP - 83

EP - 86

JO - FEBS Letters

JF - FEBS Letters

SN - 0014-5793

IS - 1-2

ER -