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Expression, purification and crystallization of GSK3β in complex with the flavonoid morin
Jeong Seok Cha,Kuglae Kim,Jinsook Ahn,Hyun-Soo Cho 한국구조생물학회 2018 Biodesign Vol.6 No.3
GSK3β is an important kinase that functions in cellular signaling pathways. Morin, a flavonoid that is plentiful in nature, was found as an inhibitor of GSK3β that can reduce tau pathology in vivo and in vitro. To identify how morin inhibits GSK3β, GSK3β protein was overexpressed and purified using affinity and ion exchange chromatography. GSK3β protein was crystallized with morin using hanging drop vapor diffusion method in the presence of 18 % (v/v) PEG 4000, 100 mM sodium citrate (pH 6.5), and 5 % (v/v) 2-propanol at 290 K. X-ray diffraction data was collected to a maximum resolution of 2.14 Å. The crystal belonged to P2 1 , with unit cell parameters a = 67.6 Å, b = 134.4 Å, c = 100.4 Å, α = γ = 90.0°, β = 103.8°.
( Wonjun Choi ),( Jinsook Seok ),( Inyoung Choi ),( Jikwon Park ),( Jeongkyu Shin ),( Soonae Lee ),( Wonyoung Paik ),( Jonghak Lee ) 대한산부인과학회 2015 Obstetrics & Gynecology Science Vol.58 No.5
Revascularization is critical for successful ovarian tissue transplantation. Vascular endothelial growth factor (VEGF) and angiopoietin-2 (angpt-2) are the principal mediators of neovascularization. This study was designed to assess VEGF and angpt-2 levels in cryopreserved ovarian tissue after heterotopic autotransplantation.Ovarian tissues harvested from ICR mice at 5 to 6 weeks of age were stratified as follows: no cryopreservation (controls, group I); vitrification in VFS-40 (vitrification, group II); and gradual freezing in dimethyl sulfoxide (slow-freezing, group III). Frozen specimens were thawed at room temperature, assaying VEGF and angpt-2 levels 1 week after cryopreservation and 2 weeks after autotransplantation.VEGF and angpt-2 protein levels were significantly lower in cryopreserved ovaries of groups II and III than in controls (group I, P<0.05), whereas groups II and III did not differ significantly in this regard. After autotransplantation of cryopreserved ovarian tissue, VEGF and angpt-2 protein levels did not differ significantly by technique but tended to be lower than corresponding levels in controls.Expression of angiogenic factors in ovarian tissue is thought to vary by method of cryopreservation. Our findings indicate that levels of angiogenic factors expressed in cryopreserved ovarian tissue after autotransplantation do not differ appreciably from control levels, regardless of cryopreservation technique.
Crystal structure of GSK3β in complex with the flavonoid, morin
Kim, Kuglae,Cha, Jeong Seok,Kim, Jin-Sik,Ahn, Jinsook,Ha, Nam-Chul,Cho, Hyun-Soo Elsevier 2018 Biochemical and biophysical research communication Vol.504 No.2
<P><B>Abstract</B></P> <P>GSK3β is a key kinase that plays a role in cellular signaling pathways. In Alzheimer's disease (AD), GSK3β has been implicated in hyperphosphorylation of tau proteins in the neuron, which is a hallmark of AD. Morin, a flavonoid that is abundant in nature, was found as an inhibitor of GSK3β that can reduce tau pathology <I>in vivo</I> and <I>in vitro</I>. In this study, we determined the crystal structure of GSK3β in complex with morin. The structure revealed that morin inhibits GSK3β by binding to the ATP binding pocket. Our findings augment the potential of morin as a functional food to help prevent AD, as well as to provide structural information to develop new therapeutics based on the morin skeleton.</P> <P><B>Graphical abstract</B></P> <P>[DISPLAY OMISSION]</P>