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논문 기본 정보
- 자료유형
- 학술저널
- 저자정보
- 발행연도
- 2010.1
- 수록면
- 541 - 546 (6page)
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We utilized a mammalian expression system to purify and characterize autotaxin (ATX)/lysophospholipase D, an enzyme present in the blood responsible for biosynthesis of lysophosphatidic acid. The human ATX cDNA encoding amino acids 29-915 was cloned downstream of a secretion signal of CD5.
At the carboxyl terminus was a thrombin cleavage site followed by the constant domain (Fc) of IgG to facilitate protein purification. The ATX-Fc fusion protein was expressed in HEK293 cells and isolated from conditioned medium of a stable clone by affinity chromatography with Protein A sepharose followed by cleavage with thrombin. The untagged ATX protein was further purified to essential homogeneity by gel filtration chromatography with a yield of approximately 5 mg/liter medium. The purified ATX protein was enzymatically active and biologically functional, offering a useful tool for further biological and structural studies of this important enzyme.
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참고문헌 (20)
참고문헌 신청
Barton, W. A. / 2006 / Crystal structures of the Tie2 receptor ectodomain and the angiopoietin-2-Tie2 complex / Nat. Struc. Mol. Biol. 13 : 524 ~ 532
Bollen, M. / 2000 / Nucleotide pyrophosphatases/phosphodiesterases on the move / Critical Rev. Biochem. Mol. Biol. 35 : 393 ~ 432
Fang, X. / 2004 / Mechanisms for lysophosphatidic acidinduced cytokine production in ovarian cancer cells / J. Biol. Chem. 279 : 9653 ~ 9661
Ferry, G. / 2003 / Autotaxin is released from adipocytes, catalyzes lysophosphatidic acid synthesis, and activates preadipocyte proliferation. Upregulated expression with ad
Giganti, A. / 2008 / Murine and Human Autotaxin α, β, and γ Isoforms: gene organization, tissue distribution, and biochemical characterization / J. Biol. Chem. 283 : 7776
Bollen, M. / 2000 / Nucleotide pyrophosphatases/phosphodiesterases on the move / Critical Rev. Biochem. Mol. Biol. 35 : 393 ~ 432
Fang, X. / 2004 / Mechanisms for lysophosphatidic acidinduced cytokine production in ovarian cancer cells / J. Biol. Chem. 279 : 9653 ~ 9661
Ferry, G. / 2003 / Autotaxin is released from adipocytes, catalyzes lysophosphatidic acid synthesis, and activates preadipocyte proliferation. Upregulated expression with ad
Giganti, A. / 2008 / Murine and Human Autotaxin α, β, and γ Isoforms: gene organization, tissue distribution, and biochemical characterization / J. Biol. Chem. 283 : 7776
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