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

FEBS Letters

Ariga, Toshio¹  Yu, Robert K.¹  Gu, Tian-Jue¹  Gu, Xin-Bin¹ 

[1] Department of Biochemistry and Molecular Biophysics, Virginia Commonwealth University, Medical College of Virginia, Richmond, VA 23298-0614, USA

关键词: Glycosyltransferase;    Sialyltransferase;    Glycolipid;    Ganglioside;    GM1;    NeuAc;    N-acetylneuraminic acid;    CMP;    cytidine monophosphate;   

DOI  :  10.1016/0014-5793(90)81444-S

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

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 M _r 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 K _m value for GM1 was 7.5 × 10⁻² M, and for CMP-NeuAc it was 6.5 × 10⁻⁵ M.

【 授权许可】

Unknown