First Synthetic Probe for the Detection and Quantification of a Protein with a Potential α,(2→8)Sialyltransferase Activity
There is considerable interest in monitoring α,(2→8)sialyltransferase (ST8) levels; however, there are few specific and sensitive methods to directly detect and quantitate the protein. This paper reports the development of a synthetic probe composed of oxidized colominic acid coupled to biotinyl-l-l...
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| Veröffentlicht in: | Bioconjugate chemistry 2004-09, Vol.15 (5), p.1084-1087 |
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| container_title | Bioconjugate chemistry |
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| creator | D'Andrea, Gabriele Lizzi, Anna Rita Oratore, Arduino |
| description | There is considerable interest in monitoring α,(2→8)sialyltransferase (ST8) levels; however, there are few specific and sensitive methods to directly detect and quantitate the protein. This paper reports the development of a synthetic probe composed of oxidized colominic acid coupled to biotinyl-l-lysine hydrazide to detect and quantify ST8 with putative “initiase” activity and its use in three solid-phase applications. The detection limit observed for ST8 purified from K562 cells was ∼2 pg by dot-blot analysis. In Western blots the probe bound and specifically recognized a protein band corresponding to ST8. In ELISA a linear dose response was obtained for pure protein in the range of 50−200 pg. Analysis of 3‘-azido-3‘deoxythymidine-treated cells by all three methods showed a reduction in ST8 compared to control cells; treated cells had 73% of control levels by ELISA. This probe will be useful for studies on the expression ST8 and its role in glycoconjugate biosynthesis. |
| doi_str_mv | 10.1021/bc049970s |
| format | Article |
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This paper reports the development of a synthetic probe composed of oxidized colominic acid coupled to biotinyl-l-lysine hydrazide to detect and quantify ST8 with putative “initiase” activity and its use in three solid-phase applications. The detection limit observed for ST8 purified from K562 cells was ∼2 pg by dot-blot analysis. In Western blots the probe bound and specifically recognized a protein band corresponding to ST8. In ELISA a linear dose response was obtained for pure protein in the range of 50−200 pg. Analysis of 3‘-azido-3‘deoxythymidine-treated cells by all three methods showed a reduction in ST8 compared to control cells; treated cells had 73% of control levels by ELISA. 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| fulltext | fulltext |
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| ispartof | Bioconjugate chemistry, 2004-09, Vol.15 (5), p.1084-1087 |
| issn | 1043-1802 1520-4812 |
| language | eng |
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| source | MEDLINE; American Chemical Society Journals |
| subjects | Biochemistry Cell Line, Tumor Chemical synthesis Data analysis Enzymes Humans Molecular Probes - analysis Molecular Probes - chemical synthesis Polysaccharides - analysis Polysaccharides - chemical synthesis Proteins Proteins - analysis Sialyltransferases - analysis |
| title | First Synthetic Probe for the Detection and Quantification of a Protein with a Potential α,(2→8)Sialyltransferase Activity |
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