Isolation to homogeneity and partial characterization of a histo-blood group A defined Fuc alpha 1 arrow right 2Gal alpha 1 arrow right 3-N-acetylgalactosaminyl-transferase from human lung tissue

The soluble histo-blood group A glycosyltransferase (Fuc alpha 1 arrow right Gal alpha 1 arrow right 3-N-acetylgalactosaminyl-transferase) was purified approximately equals 600,000-fold to homogeneity from human lung tissue. The enzyme was solubilized in 1% Triton X-100, partially purified by affini...

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Bibliographic Details
Published in:The Journal of biological chemistry 1990-01, Vol.265 (2), p.1139-1145
Main Authors: Clausen, H, White, T, Takio, K, Titani, K, Stroud, M, Holmes, E, Karkov, J, Thim, L, Hakomori, S-I
Format: Article
Language:English
Subjects:
lung
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Summary:The soluble histo-blood group A glycosyltransferase (Fuc alpha 1 arrow right Gal alpha 1 arrow right 3-N-acetylgalactosaminyl-transferase) was purified approximately equals 600,000-fold to homogeneity from human lung tissue. The enzyme was solubilized in 1% Triton X-100, partially purified by affinity chromatography on Sepharose 4B, and eluted with UDP. Final purification was obtained by twice repeated fast protein liquid chromatography ion exchange (Mono S super(TM)) with NaCl gradient elution and reverse-phase chromatography (proRPC) with acetonitrile gradient elution. Amino acid composition and N-terminal amino acid sequence of the intact transferase, as well as of peptides released by endolysyl peptidase digest or cyanogen bromide cleavage, are presented.
ISSN:0021-9258