AIM: To isolate a novel isoform of human HPO (HPO-205) from human fetal liver Marathon-ready cDNA and characterize its primary biological function. METHODS: 5'-RACE (rapid amplification of cDNA 5' ends) was used to isolate a novel isoform of hHPO in this paper. The constructed pcDNA(HPO-205), pcDNA(HPO) and pcDNA eukaryotic expression vectors were respectively transfected by lipofectamine method and the stimulation of DNA synthesis was observed by (3)H-TdR incorporation assay. Proteins extracted from different cells were analyzed by Western blot. RESULTS: A novel isoform of hHPO (HPO-205) encoding a 205 amino acid ORF corresponding to a translated production of 23 kDa was isolated and distinguished from the previous HPO that lacked the N-terminal 80 amino acids. The dose-dependent stimulation of DNA synthesis of HepG2 hepatoma cells by HPO-205 demonstrated its similar biological activity with HPO in vitro. The level of MAPK (Mitogen-activated protein kinase) phosphorylation by Western blot analysis revealed that HPO-205 might have the stronger activity of stimulating hepatic cell proliferation than that of HPO. CONCLUSION: A novel isoform of hHPO (HPO-205) was isolated from hepatic-derived cells. The comparison of HPO-205 and HPO will lead to a new insight into the structure and function of hHPO, and provide the new way of thinking to deeply elucidate the biological roles of HPO/ALR.
AIM: To isolate a novel isoform of human HPO (HPO-205) from human fetal liver Marathon-ready cDNA and characterize its primary biological function. METHODS: 5'-RACE (rapid amplification of cDNA 5' ends) was used to isolate a novel isoform of hHPO in this paper. The constructed pcDNA(HPO-205), pcDNA(HPO) and pcDNA eukaryotic expression vectors were respectively transfected by lipofectamine method and the stimulation of DNA synthesis was observed by (3)H-TdR incorporation assay. Proteins extracted from different cells were analyzed by Western blot. RESULTS: A novel isoform of hHPO (HPO-205) encoding a 205 amino acid ORF corresponding to a translated production of 23 kDa was isolated and distinguished from the previous HPO that lacked the N-terminal 80 amino acids. The dose-dependent stimulation of DNA synthesis of HepG2 hepatoma cells by HPO-205 demonstrated its similar biological activity with HPO in vitro. The level of MAPK (Mitogen-activated protein kinase) phosphorylation by Western blot analysis revealed that HPO-205 might have the stronger activity of stimulating hepatic cell proliferation than that of HPO. CONCLUSION: A novel isoform of hHPO (HPO-205) was isolated from hepatic-derived cells. The comparison of HPO-205 and HPO will lead to a new insight into the structure and function of hHPO, and provide the new way of thinking to deeply elucidate the biological roles of HPO/ALR.
Authors: G Wang; X Yang; Y Zhang; Q Wang; H Chen; H Wei; G Xing; L Xie; Z Hu; C Zhang; D Fang; C Wu; F He Journal: J Biol Chem Date: 1999-04-23 Impact factor: 5.157
Authors: C R Gandhi; R Kuddus; V M Subbotin; J Prelich; N Murase; A S Rao; M A Nalesnik; S C Watkins; A DeLeo; M Trucco; T E Starzl Journal: Hepatology Date: 1999-05 Impact factor: 17.425
Authors: Y Li; M Li; G Xing; Z Hu; Q Wang; C Dong; H Wei; G Fan; J Chen; X Yang; S Zhao; H Chen; K Guan; C Wu; C Zhang; F He Journal: J Biol Chem Date: 2000-12-01 Impact factor: 5.157
Authors: A Francavilla; N L Vujanovic; L Polimeno; A Azzarone; A Iacobellis; A Deleo; M Hagiya; T L Whiteside; T E Starzl Journal: Hepatology Date: 1997-02 Impact factor: 17.425
Authors: Yan Li; Muhammad Farooq; Donglai Sheng; Chanchal Chandramouli; Tian Lan; Nilesh K Mahajan; R Manjunatha Kini; Yunhan Hong; Thomas Lisowsky; Ruowen Ge Journal: PLoS One Date: 2012-01-26 Impact factor: 3.240