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Translation rate underpins specific targeting of N-terminal transmembrane proteins to mitochondria

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Journal of Integrative Plant Biology 2023年第6期65卷 1505-1520页

作者： Junho lee Byeongho Moon dong wook lee Inhwan Hwang Department of Life Science Pohang University of Science and TechnologyPohang 790-784South Korea Department of Integrative Food Bioscience and BiotechnologyChonnam National UniversityGwangju 61186South Korea Department Bioenergy Science and Technology Chonnam National UniversityGwangju 61186South Korea

Protein biogenesis is a complex process,and complexity is greatly increased in eukaryotic cells through specific targeting of proteins to different *** direct targeting,organellar proteins carry an organelle-specific targeting signal for recognition by organelle-specific import ***,the situation is confusing for transmembrane domain(TMD)-containing signalanchored(SA)proteins of various organelles because TMDs function as an endoplasmic reticulum(ER)targeting *** ER targeting of SA proteins is well understood,how they are targeted to mitochondria and chloroplasts remains ***,we investigated how the targeting specificity of SA proteins is determined for specific targeting to mitochondria and *** targeting requires multiple motifs around and within TMDs:a basic residue and an arginine-rich region flanking the N-and C-termini of TMDs,respectively,and an aromatic residue in the C-terminal side of the TMD that specify mitochondrial targeting in an additive *** motifs play a role in slowing down the elongation speed during translation,thereby ensuring mitochondrial targeting in a cotranslational *** contrast,the absence of any of these motifs individually or together causes at varying degrees chloroplast targeting that occurs in a post-translational manner.

关键词： arginine-rich region ER targeting mitochondria targeting organelle targeting TMD translational slowdown transmembrane domain

Molecular Mechanism of the Specificity of Protein Import into Chloroplasts and Mitochondria in Plant Cells

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Molecular Plant 2019年第7期12卷 951-966页

作者： dong wook lee Sumin lee Junho lee Seungjin Woo Md.Abdur Razzak Alessandro Vitale Inhwan Hwang Division of Integrative Biosciences and Biotechnology Pohang University of Science and Technology Pohang 37673 Korea Department of Life Sciences Pohang University of Science and Technology Pohang 37673 Korea Istituto di Biologia e Biotecnologia Agraria Consiglio Nazionale Delle Ricerche Milano Italy

Plants possess both types of endosymbiotic organelles, chloroplasts and mitochondria. Transit peptides and presequences function as signal sequences for specific import into chloroplasts and mitochondria, respectively. However, how these highly similar signal sequences confer the protein import specificity remains elusive. Here, we show that mitochondrial- or chloroplast-specific import involves two distinct steps, specificity determination and translocation across envelopes, which are mediated by the N-terminal regions and functionally interchangeable C-terminal regions, respectively, of transit peptides and presequences. A domain harboring multiple-arginine and hydrophobic sequence motifs in the N-terminal regions of presequences was identified as the mitochondrial specificity factor. The presence of this domain and the absence of arginine residues in the N-terminal regions of otherwise common targeting signals confers specificity of protein import into mitochondria and chloroplasts, respectively. AtToc159, a chloroplast import receptor, also contributes to determining chloroplast import specificity. We propose that common ancestral sequences were functionalized into mitochondrial- and chloroplast-specific signal sequences by the presence and absence, respectively, of multiple-arginine and hydrophobic sequence motifs in the N-terminal region.

关键词： transit peptide presequence protein import into chloroplasts and mitochondria N-terminal specificity domain C-terminal common translocation domain import specificity determination

Understanding protein translocation across chloroplast membranes:Translocons and motor proteins

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Journal of Integrative Plant Biology 2023年第2期65卷 408-416页

作者： Da Been Kim Changhee Na Inhwan Hwang dong wook lee Department of Integrative Food Bioscience and BiotechnologyChonnam National UniversityGwangju 61186Korea Department of Life Sciences Pohang University of Science and TechnologyPohang 37673Korea Department of Bioenergy Science and Technology Chonnam National UniversityGwangju 61186Korea Kumho Life Science Laboratory Chonnam National UniversityGwangju 61186Korea

Subcellular organelles in eukaryotes are surrounded by lipid *** an endomembrane system,vesicle trafficking is the primary mechanism for the delivery of organellar proteins to specific ***,organellar proteins for chloroplasts,mitochondria,the nucleus,and peroxisomes that are translated in the cytosol are directly imported into their target *** are a plant-specific organelle with outer and inner envelope membranes,a dual-membrane structure that is similar to *** chloroplast proteins translated by cytosolic ribosomes are thus translocated through TOC and TIC complexes(translocons in the outer and inner envelope of chloroplasts,respectively),with stromal ATPase motor proteins playing a critical role in pulling pre-proteins through these import *** the last three decades,the identity and function of TOC/TIC components and stromal motor proteins have been actively investigated,which has shed light on the action mechanisms at a molecular ***,there remains some disagreement over the exact composition of TIC complexes and genuine stromal motor *** this review,we discuss recent findings on the mechanisms by which proteins are translocated through TOC/TIC complexes and discuss future prospects for this field of research.

关键词： ATPase motor protein chloroplast molecular chaperone protein translocation TOC/TIC translocon

Nanostructured hafnium-doped strontium oxide film for homeotropic/homogeneous convertible liquid crystal alignment depending on the curing temperature

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International Journal of Smart and Nano Materials 2022年第4期13卷 597-611页

作者： dong Hyun Kim dong wook lee Jin Young Oh Jonghoon Won Hae-Chang Jeong Dae-Shik Seo IT Nano Electronic Device Laboratory Department of Electrical and Electronic EngineeringYonsei UniversitySeoulRepublic of Korea Department of Smart Manufacturing Engineering Changwon National UniversityChangwonGyeongnamRepublic of Korea Department of Electrical Electronic and Control Engineering Changwon National UniversityChangwonGyeongnamRepublic of Korea

A hafnium strontium oxide(HfSrO)liquid crystal(LC)alignment film was efficiently created through brush coating,and its ability to change the LC alignment direction was confirmed.A brush was applied to HfSrO sol coated on an indium-tin oxide substrate,after which the coating was solidified at var-ious curing *** was confirmed that a directional micro/nanostructure was formed above 280°C due to the shear stresses caused by the movement of the brush *** chemical changes were analyzed by using X-ray photoelectron spectroscopy and contact angle *** the curing temperature increased,the prevalence of oxygen bonds increased and the contact angle decreased,thereby increasing the surface *** anisotropic boundary of the micro-grooves and the van der Waals forces due to an increase in surface energy changed the alignment direction of LC mole-cules from vertical to horizontal,as verified through polarized optical microscopy and pretilt angle ***,the efficiency of the brush-coating method,which dramatically simplifies the LC alignment film process,was *** homeotropic/homogeneous LC alignment property of the HfSrO film produced through brush coating depending on the curing temperature provides an innovative approach for LC alignment.

关键词： Brush coating nanostructure solution process liquid crystal alignment

Arabidopsis BAG1 Functions as a Cofactor in Hsc70-Mediated Proteasomal Degradation of Unimported Plastid Proteins

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Molecular Plant 2016年第10期9卷 1428-1431页

作者： dong wook lee Su Jin Kim Young Jun Oh Bongsoo Choi Juhun lee Inhwan Hwang Division of Integrative Biosciences and Biotechnology Pohang University of Science and Technology Pohang 790-784 Korea

Dear Editor,Eliminating misfolded or mistargeted proteins is crucial for cell viability because these proteins accumulate as non-specific aggregates, which can be toxic to the cell （lee et al., 2009; Sroka et al., 2009）. Previously, we have shown that in ppi2 （plastid protein import 2） mutant plants, the transcript levels of Hsc70-4 （one isoform of the Hsc70 family） and CHIP （an E3 ligase） were highly upregulated, which ultimately plays crucial roles in proteasomal degradation of unimported plastid proteins （lee et al., 2009）. We also found that, along with those of Hsc70-4 and CHIP, the transcript level of AtBAG1 （Arabidopsis thaliana Bcl2-associated athanogene 1） in the ppi2 mutant was 2.38-fold higher than that in the wild-type （lee et al., 2009）.

关键词： SEQUENCE MOTIFS

Cytochrome b5 Reductase 1 Triggers Serial Reactions that Lead to Iron Uptake in Plants

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Molecular Plant 2016年第4期9卷 501-513页

作者： Young Jun Oh Hanul Kim Sung Hee Seo Bae Geun Hwang Yoon Seok Chang Junho lee dong wook lee Eun Ju Sohn Sang Joon lee Youngsook lee Inhwan Hwang Division of Integrative Biosciences and Biotechnology Department Life Sciences Division of Environmental Science and Engineering Division of Mechanical Engineering Pohang University of Science and Technology Pohang 790-784 Korea

Rhizosphere acidification is essential for iron （Fe） uptake into plant roots. Plasma membrane （PM） H＊-ATPases play key roles in rhizosphere acidification. However, it is not fully understood how PM H＋-ATPase activity is regulated to enhance root Fe uptake under Fe-deficient conditions. Here, we present evidence that cytochrome b5 reductase 1 （CBR1） increases the levels of unsaturated fatty acids, which stimulate PM H＋-ATPase activity and thus lead to rhizosphere acidification. CBRl-overexpressing （CBRI-OX） Arabidopsis thaliana plants had higher levels of unsaturated fatty acids （18：2 and 18：3）, higher PM H＊-ATPase activity, and lower rhizosphere pH than wild-type plants. By contrast, cbrl loss-of-function mutant plants showed lower levels of unsaturated fatty acids and lower PM H＊-ATPase activity but higher rhizosphere pH. Reduced PM H＊-ATPase activity in cbrl could be restored in vitro by addition of unsatu- rated fatty acids. Transcript levels of CBR1, fatty acids desaturase 2 （FAD2）, and fatty acids desaturase 3 （FAD3） were increased under Fe-deficient conditions. We propose that CBR1 has a crucial role in increasing the levels of unsaturated fatty acids, which activate the PM H＊-ATPase and thus reduce rhizosphere pH. This reaction cascade ultimately promotes root Fe uptake.

关键词： cytochrome b5 reductase 1 (CBR1) unsaturated fatty acids iron (Fe) uptake H^+-ATPase

Interactions between Transmembrane Helices within Monomers of the Aquaporin AtPIP2;1 Play a Crucial Role in Tetramer Formation

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Molecular Plant 2016年第7期9卷 1004-1017页

作者： Yun-Joo Yoo Hyun Kyung lee Wonhee Han Dae Heon Kim Myoung Hui lee Jouhyun Jeon dong wook lee Junho lee Yongjik lee Juhun lee Jin Seok Kim Yunje Cho Jin-Kwan Han Inhwan Hwang School of Interdisciplinary Bioscience and Bioengineering Pohang University of Science and Technology Pohang 790-784 Korea Department of Life Sciences Pohang University of Science and Technology Pohang 790-784 Korea Department of Biology Sunchon National University Sunchon 57922 Korea Division of Integrative Biosciences and Biotechnology Pohang University of Science and Technology Pohang 790-784 Korea

Aquaporin （AQP） is a water channel protein found in various subcellular membranes of both prokaryotic and eukaryotic cells. The physiological functions of AQPs have been elucidated in many organisms. However, understanding their biogenesis remains elusive, particularly regarding how they assemble into tetramers. Here, we investigated the amino acid residues involved in the tetramer formation of the Arabidopsis plasma membrane AQP AtPIP2;1 using extensive amino acid substitution mutagenesis. The mutant proteins V41A/ E44A, F51A/L52A, F87A/191A, F92A/193A, V95A/Y96A, and H216A/L217A, harboring alanine substitutions in the transmembrane （TM） helices of AtPIP2;1 polymerized into multiple oligomeric complexes with a vari- able number of subunits greater than four. Moreover, these mutant proteins failed to traffic to the plasma membrane, instead of accumulating in the endoplasmic reticulum （ER）. Structure-based modeling revealed that these residues are largely involved in interactions between TM helices within monomers. These results suggest that inter-TM interactions occurring both within and between monomers play crucial roles in tetramer formation in the AtPIP2;1 complex. Moreover, the assembly of AtPIP2;1 tetramers is critical for their trafficking from the ER to the plasma membrane, as well as water permeability.

关键词： aquaporin AtPIP2 tetramer formation interaction between transmembrane helices

Incorporation of Ionic Conductive Polymers into Sulfide Electrolyte-Based Solid-State Batteries to Enhance Electrochemical Stability and Cycle Life

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Energy & Environmental Materials 2024年第6期7卷 4-12页

作者： Juhyoung Kim Woonghee Choi Seong-Ju Hwang dong wook Kim Advanced Materials Division Korea Research Institute of Chemical Technology(KRICT)Daejeon 34114Korea Department of Material Science and Engineering Yonsei UniversitySeoul 03722Korea Department of Material Science and Engineering Seoul National UniversitySeoul 08826Korea Advanced Materials and Chemical Engineering University of Science and Technology(UST)Daejeon 34113Korea

Sulfide-based inorganic solid electrolytes are promising materials for high-performance safe solid-state *** high ion conductivity,mechanical characteristics,and good processability of sulfide-based inorganic solid electrolytes are desirable properties for realizing high-performance safe solid-state batteries by replacing conventional liquid ***,the low chemical and electrochemical stability of sulfide-based inorganic solid electrolytes hinder the commercialization of sulfide-based safe solid-state ***,the instability of sulfide-based inorganic solid electrolytes is intensified in the cathode,comprising various *** this study,carbonate-based ionic conductive polymers are introduced to the cathode to protect cathode materials and suppress the reactivity of sulfide *** instruments,including electrochemical spectroscopy,X-ray photoelectron spectroscopy,and scanning electron microscopy,confirm the chemical and electrochemical stability of the polymer electrolytes in contact with sulfide-based inorganic solid ***-based solid-state cells show stable electrochemical performance over 100 cycles when the ionic conductive polymers were applied to the cathode.

关键词： composite cathode electrochemical stability ionic conductive polymer solid-state battery sulfide solid electrolyte

Highly Aligned Ternary Nanofiber Matrices Loaded with MXene Expedite Regeneration of Volumetric Muscle Loss

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Nano-Micro Letters 2024年第4期16卷 269-292页

作者： Moon Sung Kang Yeuni Yu Rowoon Park Hye Jin Heo Seok Hyun lee Suck Won Hong Yun Hak Kim dong‑wook Han Department of Cogno‑Mechatronics Engineering College of Nanoscience and NanotechnologyPusan National UniversityBusan 46241Republic of Korea Medical Research Institute School of MedicinePusan National UniversityYangsan 50612Republic of Korea Department of Anatomy School of MedicinePusan National UniversityYangsan 50612Republic of Korea Engineering Research Center for Color‑Modulated Extra‑Sensory Perception Technology Pusan National UniversityBusan 46241Republic of Korea Department of Biomedical Informatics School of MedicinePusan National UniversityYangsan 50612Republic of Korea Periodontal Disease Signaling Network Research Center and Dental and Life Science Institute School of DentistryPusan National UniversityYangsan 50612Republic of Korea BIO‑IT Fusion Technology Research Institute Pusan National UniversityBusan 46241Republic of Korea Present Address:Osstem Implant Inc. Seoul 07789Republic of Korea

Current therapeutic approaches for volumetric muscle loss(VML)face challenges due to limited graft availability and insufficient *** overcome these limitations,tissue-engineered scaffolds have emerged as a promising *** this study,we developed aligned ternary nanofibrous matrices comprised of poly(lactide-co-ε-caprolactone)integrated with collagen and Ti_(3)C_(2)T_(x)MXene nanoparticles(NPs)(PCM matrices),and explored their myogenic potential for skeletal muscle tissue *** PCM matrices demonstrated favorable physicochemical properties,including structural uniformity,alignment,microporosity,and *** vitro assays revealed that the PCM matrices promoted cellular behaviors and myogenic differentiation of C2C12 ***,in vivo experiments demonstrated enhanced muscle remodeling and recovery in mice treated with PCM matrices following VML *** insights from next-generation sequencing revealed that MXene NPs facilitated protein and ion availability within PCM matrices,leading to elevated intracellular Ca^(2+)levels in myoblasts through the activation of inducible nitric oxide synthase(i NOS)and serum/glucocorticoid regulated kinase 1(SGK1),ultimately promoting myogenic differentiation via the m TOR-AKT ***,upregulated i NOS and increased NO–contributed to myoblast proliferation and fiber fusion,thereby facilitating overall myoblast *** findings underscore the potential of MXene NPs loaded within highly aligned matrices as therapeutic agents to promote skeletal muscle tissue recovery.

关键词： Ti_(3)C_(2)T_(x)MXene nanoparticle Ternary nanofibrous matrices Myogenesis Regeneration of volumetric muscle loss Next generation sequencing