Human PEP-1-ribosomal protein S3 protects against UV-induced skin cell death

Choi, Soo Hyun,Kim, So Young,An, Jae Jin,Lee, Sun Hwa,Kim, Dae Won,Ryu, Hea Jin,Lee, Nam Il,Yeo, Seung Il,Jang, Sang Ho,Won, Moo Ho,Kang, Tae-Cheon,Kwon, Hyung Joo,Cho, Sung-Woo,Kim, Joon,Lee, Kil Soo Elsevier 2006 FEBS letters Vol.580 No.30

<P><B>Abstract</B></P><P>The consequences of ultraviolet (UV) exposure are implicated in skin aging and cell death. The ribosomal protein S3 (rpS3) is one of the major proteins by which cells counteract the deleterious effects of UV and it plays a role in the repair of damaged DNA. In the present study, we investigated the protective effects of PEP-1-rpS3 fusion protein after UV-induced cell injury. A human rpS3 gene was fused with PEP-1 peptide in a bacterial expression vector to produce a genetic in-frame PEP-1-rpS3 fusion protein. The expressed and purified fusion proteins were efficiently transduced into skin cells in a time- and dose-dependent manner. Once inside the cells, transduced PEP-1-rpS3 fusion protein was stable for 48h. We showed that transduced PEP-1-rpS3 fusion protein increased cell viability and dramatically reduced DNA lesions in the UV exposed skin cells. Immunohistochemical analysis revealed that PEP-1-rpS3 fusion protein efficiently penetrated the epidermis as well as the dermis of the subcutaneous layer when sprayed on animal skin. These results suggest that PEP-1-rpS3 fusion protein can be used in protein therapy for various disorders related to UV, including skin aging and cancer.</P>

Protein nanoparticles directed cancer imaging and therapy

Miao Yao,Yang Tao,Yang Shuxu,Yang Mingying,Mao Chuanbin 나노기술연구협의회 2022 Nano Convergence Vol.9 No.2

Cancer has been a serious threat to human health. Among drug delivery carriers, protein nanoparticles are unique because of their mild and environmentally friendly preparation methods. They also inherit desired characteristics from natural proteins, such as biocompatibility and biodegradability. Therefore, they have solved some problems inherent to inorganic nanocarriers such as poor biocompatibility. Also, the surface groups and cavity of protein nanoparticles allow for easy surface modification and drug loading. Besides, protein nanoparticles can be combined with inorganic nanoparticles or contrast agents to form multifunctional theranostic platforms. This review introduces representative protein nanoparticles applicable in cancer theranostics, including virus-like particles, albumin nanoparticles, silk protein nanoparticles, and ferritin nanoparticles. It also describes the common methods for preparing them. It then critically analyzes the use of a variety of protein nanoparticles in improved cancer imaging and therapy.

Transduced Tat-DJ-1 protein inhibits cytokines-induced pancreatic RINm5F cell death

( Hyo Sang Jo ),( Hyeon Ji Yeo ),( Hyun Ju Cha ),( Sang Jin Kim ),( Su Bin Cho ),( Jung Hwan Park ),( Chi Hern Lee ),( Eun Ji Yeo ),( Yeon Joo Choi ),( Won Sik Eum ),( Soo Young Choi ) 생화학분자생물학회(구 한국생화학분자생물학회) 2016 BMB Reports Vol.49 No.5

Loss of pancreatic β-cells by oxidative stress or cytokines is associated with diabetes mellitus (DM). DJ-1 is known to as a multifunctional protein, which plays an important role in cell survival. We prepared cell permeable wild type (WT) and mutant type (M26I) Tat-DJ-1 proteins to investigate the effects of DJ-1 against combined cytokines (IL-1β, IFN-γ and TNF-α)-induced RINm5F cell death. Both Tat-DJ-1 proteins were transduced into RINm5F cells. WT Tat-DJ-1 proteins significantly protected against cell death from cytokines by reducing intracellular toxicities. Also, WT Tat-DJ-1 proteins markedly regulated cytokines-induced pro- and anti-apoptosis proteins. However, M26I Tat-DJ-1 protein showed relatively low protective effects, as compared to WT Tat-DJ-1 protein. Our experiments demonstrated that WT Tat-DJ-1 protein protects against cytokine-induced RINm5F cell death by suppressing intracellular toxicities and regulating apoptosisrelated protein expression. Thus, WT Tat-DJ-1 protein could potentially serve as a therapeutic agent for DM and cytokine related diseases. [BMB Reports 2016; 49(5): 297-302]

Tat-CIAPIN1 protein prevents against cytokine-induced cytotoxicity in pancreatic RINm5Fβ-cells

( Hyeon Ji Yeo ),( Min Jea Shin ),( Dae Won Kim ),( Hyeok Yil Kwon ),( Won Sik Eum ),( Soo Young Choi ) 생화학분자생물학회 2021 BMB Reports Vol.54 No.9

Cytokines activate inflammatory signals and are major mediators in progressiveβ-cell damage, which leads to type 1 diabetes mellitus. We recently showed that the cell-permeable Tat-CIAPIN1 fusion protein inhibits neuronal cell death induced by oxidative stress. However, how the Tat-CIAPIN1 protein affects cytokine- inducedβ-cell damage has not been investigated yet. Thus, we assessed whether the Tat-CIAPIN1 protein can protect RINm5Fβ-cells against cytokine-induced cytotoxicity. In cytokineexposed RINm5Fβ-cells, the transduced Tat-CIAPIN1 protein elevated cell survivals and reduced reactive oxygen species (ROS) and DNA fragmentation levels. The Tat-CIAPIN1 protein reduced mitogen-activated protein kinases (MAPKs) and NF-βB activation levels and elevated Bcl-2 protein, whereas Bax and cleaved Caspase-3 proteins were decreased by this fusion protein. Thus, the protection of RINm5Fβ-cells by the Tat-CIAPIN1 protein against cytokine-induced cytotoxicity can suggest that the Tat-CIAPIN1 protein might be used as a therapeutic inhibitor against RINm5Fβ-cell damage. [BMB Reports 2021; 54(9): 458-463]

Transduced Tat-DJ-1 protein inhibits cytokines-induced pancreatic RINm5F cell death

( Hyo Sang Jo ),( Hyeon Ji Yeo ),( Hyun Ju Cha ),( Sang Jin Kim ),( Su Bin Cho ),( Jung Hwan Park ),( Chi Hern Lee ),( Eun Ji Yeo ),( Yeon Joo Choi ),( Won Sik Eum ),( Soo Young Choi ) 생화학분자생물학회(구 한국생화학분자생물학회) 2016 BMB Reports Vol.49 No.6

Loss of pancreatic β-cells by oxidative stress or cytokines is associated with diabetes mellitus (DM). DJ-1 is known to as a multifunctional protein, which plays an important role in cell survival. We prepared cell permeable wild type (WT) and mutant type (M26I) Tat-DJ-1 proteins to investigate the effects of DJ-1 against combined cytokines (IL-1β, IFN-Υ and TNF-α)-induced RINm5F cell death. Both Tat-DJ-1 proteins were transduced into RINm5F cells. WT Tat-DJ-1 proteins significantly protected against cell death from cytokines by reducing intracellular toxicities. Also, WT Tat-DJ-1 proteins markedly regulated cytokines-induced pro- and anti-apoptosis proteins. However, M26I Tat-DJ-1 protein showed relatively low protective effects, as compared to WT Tat-DJ-1 protein. Our experiments demonstrated that WT Tat-DJ-1 protein protects against cytokine-induced RINm5F cell death by suppressing intracellular toxicities and regulating apoptosisrelated protein expression. Thus, WT Tat-DJ-1 protein could potentially serve as a therapeutic agent for DM and cytokine related diseases. [BMB Reports 2016; 49(5): 297-302]

Membrane transducing activity of recombinant Hoxc8 proteinand its possible application as a gene delivery vector

Eun Shin Kim,박형우,김명희,강명모 생화학분자생물학회 2008 Experimental and molecular medicine Vol.40 No.2

In order to examine whether the Hoxc8 protein can deliver nucleic acid into mammalian cells, we designed several Hoxc8-derived recombinant proteins to be synthesized as glutathione S-transferase (GST) fused forms in E.coli (GST-Hoxc81-242, containing a full length of Hoxc8; GST-Hoxc8152-242, possessing a deletion of the acidic N-terminus of Hoxc8; GST-Hoxc8149-208, which contained the homeodomain only). After labeling these proteins with Oregon 488, we examined their membrane transduction ability under the fluorescence microscope and verified that all three proteins showed similar transduction efficiency. The ability of the proteins to form in vitro protein-DNA complexes was analyzed on agarose gel; both GST-Hoxc81-242 and GSTHoxc8149- 208 formed complexes. In contrast, the GSTHoxc8152- 242 protein did not form a complex. The GST-Hoxc8149-208 protein formed a complex with DNA at a mass ratio of 1:1 (DNA:protein), and GSTHoxc81- 242 formed a complex at a mass ratio of 1:5. When the DNA (pDsRed1-C1) and protein complexes were added to culture media containing mammalian cells, the cells uptook the complexes, which was indicated by red fluorescence expression under the fluorescent microscope. These results indicate that recombinant Hoxc8 derivatives that harbor a homeodomain are able to traverse the mammalian cellular membrane. DNA that is bound to the recombinant derivatives can be carried across the membrane as well. This process could be applied in the development of a useful delivery vector for gene therapy in the future.

Genetically engineered and self-assembled oncolytic protein nanoparticles for targeted cancer therapy

Lee, Joong-jae,Kang, Jung Ae,Ryu, Yiseul,Han, Sang-Soo,Nam, You Ree,Rho, Jong Kook,Choi, Dae Seong,Kang, Sun-Woong,Lee, Dong-Eun,Kim, Hak-Sung IPC Science and Technology Press 2017 Biomaterials Vol.120 No.-

<P><B>Abstract</B></P> <P>The integration of a targeted delivery with a tumour-selective agent has been considered an ideal platform for achieving high therapeutic efficacy and negligible side effects in cancer therapy. Here, we present engineered protein nanoparticles comprising a tumour-selective oncolytic protein and a targeting moiety as a new format for the targeted cancer therapy. Apoptin from chicken anaemia virus (CAV) was used as a tumour-selective apoptotic protein. An EGFR-specific repebody, which is composed of LRR (Leucine-rich repeat) modules, was employed to play a dual role as a tumour-targeting moiety and a fusion partner for producing apoptin nanoparticles in <I>E. coli</I>, respectively. The repebody was genetically fused to apoptin, and the resulting fusion protein was shown to self-assemble into supramolecular repebody-apoptin nanoparticles with high homogeneity and stability as a soluble form when expressed in <I>E. coli</I>. The repebody-apoptin nanoparticles showed a remarkable anti-tumour activity with negligible side effects in xenograft mice through a cooperative action of the two protein components with distinct functional roles. The repebody-apoptin nanoparticles can be developed as a systemic injectable and tumour-selective therapeutic protein for targeted cancer treatment.</P> <P><B>Graphical abstract</B></P> <P>[DISPLAY OMISSION]</P>

Transduced PEP-1-Grb7 Fusion Protein Suppressed LPS-induced COX-2 Expression

An, Jae-Jin,Kim, So-Young,Lee, Sun-Hwa,Kim, Dae-Won,Ryu, Hea-Jin,Yeo, Seung-Il,Jang, Sang-Ho,Kwon, Hyung-Joo,Kim, Tae-Yoon,Lee, Sang-Chul,Poo, Ha-Ryoung,Cho, Sung-Woo,Lee, Kil-Soo,Park, Jin-Seu,Eum, W Korean Society for Biochemistry and Molecular Biol 2007 Journal of biochemistry and molecular biology Vol.40 No.2

Although the incidence and severity of atopic dermatitis (AD) is steadily increasing at an alarming rate, its pathogenic mechanisms remain poorly understood yet. Recently, we found that the expression of Grb7 protein was markedly decreased in AD patients using proteomic analysis. In the present study, human Grb7 gene was fused with PEP-1 peptide in a bacterial expression vector to produce a genetic in-frame PEP-1-Grb7 fusion protein. The expressed and purified PEP-1-Grb7 fusion proteins transduced efficiently into skin cells in a time- and dose-dependent manner when added exogenously in culture media. Once inside the cells, the transduced PEP-1-Grb7 protein was stable for 48 h. In addition, transduced PEP-1-Grb7 fusion protein markedly increased cell viability in macrophage RAW 264.7 cells treated with LPS by inhibition of the COX-2 expression level. These results suggest that the PEP-1-Grb7 fusion protein can be used in protein therapy for inflammatory skin disorders, including AD.

Ischemic heart diseases: Current treatments and future

Choi, Donghoon,Hwang, Ki-Chul,Lee, Kuen-Yong,Kim, Yong-Hee Elsevier 2009 Journal of controlled release Vol.140 No.3

<P><B>Abstract</B></P><P>Ischemic heart disease is a rapidly increasing common cause of death in the world. This disease is the insufficient status of oxygen within the cardiac muscles due to an imbalance between oxygen supply and demand, and a cardiac disease that occurs as a result of coronary artery stenosis. Conventional surgery-based therapy for the treatment of ischemic heart diseases has been advanced with biopharmaceutical-based therapy, such as protein, gene and cell therapy. The conventional medical therapy focuses on the use of drug eluting stents, coronary-artery bypass-graft surgery and anti-thrombosis. Biopharmaceutical-based therapies including recombinant protein therapy, gene therapy and cell transplantation are recognized as promising approaches in inducing neovascularization and improving collateral blood flow in the ischemic heart. This review explores the current status and future of the treatment of ischemic heart diseases with conventional medical therapy, biopharmaceutical-based therapy focused on the proteins and polymeric hydrogels for delivery of therapeutic proteins.</P> <P><B>Graphical abstract</B></P><P><ce:figure></ce:figure></P>

Endothelin-1-유도 근수축에 관여하는 부활효소의 활성과 물리치료의 상관성

김미선 ( Mi-sun Kim ),김일현 ( Il-hyun Kim ),황병용 ( Byong-yong Hwang ),김중환 ( Jung-hwan Kim ) 대한물리치료학회 2008 대한물리치료학회지 Vol.20 No.3

Purpose: The non-receptor-type protein tyrosine kinase Syk (636 amino acids, 72 kDa) is ubiquitously expressed in hematopoietic stem cells and has been widely studied as a regulator and effector of B cell receptor signaling that occurs in processes such as differentiation, proliferation and apoptosis. However, the mechanism relating Syk and p38 mitogen-activated protein kinases (p38MAPK) by endothelin-1 (ET-1, 21 amino acids) stimulation in muscle cells, especially in the volume-dependent hypertensive state, remains unclear. Methods: In this study, we investigated the relationship between Syk and p38MAPK for isometric contraction and enzymatic activity by ET-1 from rat aortic smooth muscle cells and aldosterone-analogue deoxycorticosterone acetate (DOCA) hypertensive state rats (ADHR). Results: The systolic blood pressure was significantly increased in ADHR than in a control group of animals. ET-1 induced isometric contraction and phosphorylation of p38MAPK, which was increased in muscle strips from ADHR. Increased vasoconstriction and phosphorylation of p38MAPK induced by treatment with 30 nM ET-1 were inhibited by the use of 10μM SB203580, an inhibitor of p38MAPK from ADHR. Furthermore, ET-1 induced isometric contraction and phosphorylation of Syk and p38MAPK, which were increased in the aortic smooth muscle cells. Increased tension and phosphorylation of Syk and p38MAPK induced by ET-1 were inhibited by SB203580 from rat aortic smooth muscle cells. Conclusion: These results, suggest that the Syk activity affects ET-1-induced contraction through p38MAPK in smooth muscle cells and that the same pathway directly or indirectly is associated with volume dependent hypertension. The findings suggest the need to develop cardiovascular disease-specialized physical therapy.