Sp1 was first detected in HeLa cells on the basis of its ability to activate the SV40 early promoter transcription (1,2). Subsequently it was shown to recognize and bind selectively to a GC-rich consensus sequence (GC-box: GGGCGG or CACCC) that presents in the promoter of several important cellular genes, including SV40 early, HIV-1, PDGF-B etc. Sp1 was the first transcription factor to be cloned and (3). Analysis of structure and function has revealed that Sp1 can be separated into discrete functional domains. The DNA-binding domain consists of three zinc fingers that specifically bind to the GC-box element (4,5). Sp1 contains at least four separate transcriptional activation domains. Two of these domains are glutamine-rich, a well-characterized motif found in several other transcription factors (6). In addition to transcription, Sp1 function has been linked to cell growth, cancer, Huntington disease and other disorders through transcriptional regulation or specific protein-protein interactions (7-9). The function of Sp1 can be regulated by phosphorylation and glycosylation (10, 11).
GST-Sp1 was isolated from an E. coli strain that carries the coding sequence of the fused protein under the control of a T7 promoter. GST-Sp1 has been applied in protein-protein interactions assays. The GST-Sp1 fusion protein is greater than 95% homogeneous based on SDS-PAGE analysis.
1 unit equals 1 nanogram of purified protein. 100 units (ng) are sufficient for protein-protein interaction assay. Variable in different lots 1x dilution buffer A: 20 mM Tris-Cl (pH 8.0), 20% Glycerol, 100 mM KCl, 1 mM DTT and 0.2 mM EDTA
References: 1. Dynan, WS. et al., (1983) Cell 35, 79-87 2. Briggs, MR. et al., (1986) Science 234, 47-52 3. Kadonaga, JT. et al., (1987) Cell 51, 1079-1090 4. Pascal, E. et al., (1991) Genes & Dev. 5, 1646-1656 5. Nagaoka, M et al., (2001) Nucleic Acids Res. 29, 4920-4929 6. Tanaka, M. et al., (1990) Cell 60, 375-386 7. Black, AR. et al., (2001) J Cell. Physiol. 188, 143-160 8. Rafty, LA. et al., (2002) J Cell Biochem 85, 490-495 9. Dunah, AW. et al., (2002) Science 0, 10726131-0. 10. Leggett, RW. et al., (1995) J Biol. Chem. 270, 25879-25884 11. Han, I. et al., (1997) Mol. Cell. Biol. 17, 2550-2558
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