Purification and characterization of avian glycolipid: beta-galactosyltransferases (GalT-4 and GalT-3): cloning and expression of truncated betaGalT-4

• Autorzy: Basu S. S. , Dastgheib S. , Ghosh S. , Basu M. , Kelly P. , Basu S.
• Języki publikacji: EN

Abstrakty

EN

Acidic glycolipid of ganglio-(containing sialic acid) and sialyl-lactofucosyl-type, SA-Lex (containing sialic acid and fucose) are developmentally regulated and appear to be ubiqitous on neuronal and cancer cell surfaces of animals. Two glycolipid: β-galactosyltransferases, GalT-3 and GalT-4, were characterized in embryonic chicken brain (ECB). Based on substrate competition experiments, these two activities were believed to be due to expression of two gene products. A cDNA fragments (about 600 bp) encoding the catalytic domain of the GalT-4 (UDP-Gal:LcOse3Cer β1,4galactosyltransferase) from ECB and human Colo-205 were isolated. These cDNAs were expressed as a soluble glutathione-S-transferase fusion protein (48 kDa) in Eschericchia coli. Interactions between GlcNAc-, UDP-hexanolamine-, and α-lactalbumin were studied with the purified fusion protein (recombinant and truncated). Functionally it was similar to that of native GalT-4 purified (40000-fold) from 11-day-old ECB. GalT-3 (UDP-Gal:GM2β1,3galactosyltransferase) was purified from 19-day-old ECB, and a polyclonal antibody was produced against the peptide backbone for immunoscreening of a λZAP ECB cDNA expression library. Each of the GalT-3 peptides (62 and 65 kDa was analyzed by protein fingerprinting analysis indicating a similar peptide mapping pattern.

Słowa kluczowe

EN

cancer cell; glycosyltransferase; brain; animal; biosynthesis; ganglioside; galactosyltransferase; sialic acid; neuronal cell; acidic glycolipid; cloning; Escherichia coli; cDNA; chicken; beta-galactosyltransferase; glycolipid

• Wydawca: -
• Czasopismo: Acta Biochimica Polonica
• Rocznik: 1998
• Tom: 45
• Numer: 2
• Opis fizyczny: p.451-467,fig.

Twórcy

autor

Basu S. S.

• Duke University Medical School, Durham, NC, USA

autor

Dastgheib S.

autor

Ghosh S.

autor

Basu M.

autor

Kelly P.

autor

Basu S.

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