Purification and NMR studies on Phosphatase domain of UBLCP1
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UBLCP1 is composed of Ubiquitin Like domain and RNA Polymerase Ⅱ PhosphataseⅠdomain. Phosphatase domain (25.9KDa) has been cloned into the E.coli using pET32a vector with TEV protease cleavage site and successfully purified as a monomer using affinity chromatography and histidine tag was cleaved with TEV protease for structural studies. Our results indicated that the Phosphatase domain showed well-defined folded structure based on data from one-dimensional and two-dimensional NMR spectroscopy. Data form circular dichroism also suggested that Phosphatase domain consisted of both α-helix and β-sheet. This information will be used for detailed structural study of UBLCP1.
Abstract
UBLCP1 is composed of Ubiquitin Like domain and RNA Polymerase Ⅱ PhosphataseⅠdomain. Phosphatase domain (25.9KDa) has been cloned into the E.coli using pET32a vector with TEV protease cleavage site and successfully purified as a monomer using affinity chromatography and histidine tag was cleaved with TEV protease for structural studies. Our results indicated that the Phosphatase domain showed well-defined folded structure based on data from one-dimensional and two-dimensional NMR spectroscopy. Data form circular dichroism also suggested that Phosphatase domain consisted of both α-helix and β-sheet. This information will be used for detailed structural study of UBLCP1.
참고문헌 (11)
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