Evidence that high mobility group protein 17 is not phosphorylated in human colon carcinoma cells |
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Authors: | Mrunal S. Chapekar Michael Bustin Robert I. Glazer |
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Affiliation: | 1. The Laboratory of Biological Chemistry, NCI, NIH, Bethesda, MD 20205, U.S.A.;2. The Laboratory of Molecular Carcinogenesis, NCI, NIH, Bethesda, MD 20205, U.S.A. |
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Abstract: | The high mobility group proteins 14 and 17 were reported previously to be phosphorylated in murine and human tumor cell lines. Recently, it was suggested that subgroups of HMG-14, HMG-14a and 14b, but not HMG-17, were phosphorylated in situ in HeLa cells. In order to definitively determine whether HMG-17 is indeed phosphorylated or whether the protein previously identified as [32P]HMG-17 was a subgroup of HMG-14, we have used the technique of electroblotting in conjunction with an immunochemical procedure utilizing anti-HMG-17 IgG. Our results indicate that HMG-17 was not phosphorylated in human colon carcinoma cell line HT-29 incubated for 18 h with 32Pi, but that HMG-14a and HMG-14b were phosphorylated. In contrast, HMG-14a, -14b and -17 were phosphorylated in vitro in isolated nuclei incubated with [γ-32P]ATP. |
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Keywords: | High mobility group protein Phosphorylation (Human colon carcinoma) Hepes 4-(2-hydroxyethyl)-1-piperazineethane-sulfonic acid PMSF phenylmethylsulfonyl fluoride |
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