Molecular Characterization of Chicken Toll-like Receptor 7

Chai, Han-Ha,Suk, Jae Eun,Lim, Dajeong,Lee, Kyung-Tai,Choe, Changyong,Cho, Yong-Min The Korean Society of Animal Reproduction 2015 Reproductive & developmental biology Vol.39 No.4

Toll-like receptor 7 (TLR7) is critical for the triggering of innate immune response by recognizing the conserved molecular patterns of single-stranded RNA (ssRNA) viruses and mediated antigenic adaptive immunity. To understand how TLR7 distinguish pathogen-derived molecular patterns from the host self, it is essential to be able to identify TLR7 receptor interaction interfaces, such as active sites or R848-agonist binding sites. The functional interfaces of TLR7 can serve as targets for structure-based drug design in studying the TLR7 receptor's structure-function relationship. In contrast to mammalian TLR7, chicken TLR7 (chTLR7) is unknown for its important biological function. Therefore, it has been targeted to mediate contrasting evolutionary patterns of positive selection into non-synonymous SNPs across eleven species using TLR7 conservation patterns (evolutionary conserved and class-specific trace residues), where protein sequence differences to the TLR7 receptors of interest record mutation that have passed positive section across the species. In this study, we characterized the Lys609 residue on chTLR7-ECD homodimer interfaces to reflect the current tendency of evolving positive selection to be transfer into a stabilization direction of the R848-agonist/chTLR7-ECDs complex under the phylogenetically variable position across species and we suggest a potential indicator for contrasting evolutionary patterns of both the species TLR-ECDs.

Molecular Modeling of Small Molecules as BVDV RNA-Dependent RNA Polymerase Allosteric Inhibitors

Han-ha Chai,Dajeong Lim,Hee-yeoul Chai,정은경 대한화학회 2013 Bulletin of the Korean Chemical Society Vol.34 No.3

Bovine viral diarrhea virus (BVDV), a major pathogen of cattle, is a well-characterized pestivirus which has been used as a good model virus for HCV. The RNA-dependent RNA polymerase (RdRp) plays a key role in the RNA replication process, thus it has been targeted for antivirus drugs. We employed two-dimensional quantitative structure-activity relationship (2D-QSAR) and molecular field analysis (MFA) to identify the molecular substructure requirements, and the particular characteristics resulted in increased inhibitory activity for the known series of compounds to act as effective BVDV inhibitors. The 2D-QSAR study provided the rationale concept for changes in the structure to have more potent analogs focused on the class of arylazoenamines, benzimidazoles, and acridine derivatives with an optimal subset of descriptors, which have significantly contributed to overall anti-BVDV activity. MFA represented the molecular patterns responsible for the actions of antiviral compound at their receptors. We conclude that the polarity and the polarizability of a molecule play a main role in the inhibitory activity of BVDV inhibitors in the QSAR modeling.

Molecular Characterization of Chicken Toll-like Receptor 7

Han-Ha Chai,Jae Eun Suk,Dajeong Lim,Kyung-Tai Lee,Changyong Choe,Yong-Min Cho 한국동물번식학회 2015 Reproductive & developmental biology Vol.39 No.4

Toll-like receptor 7 (TLR7) is critical for the triggering of innate immune response by recognizing the conserved molecular patterns of single-stranded RNA (ssRNA) viruses and mediated antigenic adaptive immunity. To understand how TLR7 distinguish pathogen-derived molecular patterns from the host self, it is essential to be able to identify TLR7 receptor interaction interfaces, such as active sites or R848-agonist binding sites. The functional interfaces of TLR7 can serve as targets for structure-based drug design in studying the TLR7 receptor’s structure-function relationship. In contrast to mammalian TLR7, chicken TLR7 (chTLR7) is unknown for its important biological function. Therefore, it has been targeted to mediate contrasting evolutionary patterns of positive selection into non-synonymous SNPs across eleven species using TLR7 conservation patterns (evolutionary conserved and class-specific trace residues), where protein sequence differences to the TLR7 receptors of interest record mutation that have passed positive section across the species. In this study, we characterized the Lys609 residue on chTLR7-ECD homodimer interfaces to reflect the current tendency of evolving positive selection to be transfer into a stabilization direction of the R848-agonist/ chTLR7-ECDs complex under the phylogenetically variable position across species and we suggest a potential indicator for contrasting evolutionary patterns of both the species TLR-ECDs.

Molecular Characterization of Chicken Toll-like Receptor 7

Han-Ha Chai,Jae Eun Suk,Dajeong Lim,Kyung-Tai Lee,Changyong Choe,Yong-Min Cho 한국동물생명공학회(구 한국동물번식학회) 2015 Reproductive & developmental biology Vol.39 No.4

Toll-like receptor 7 (TLR7) is critical for the triggering of innate immune response by recognizing the conserved molecular patterns of single-stranded RNA (ssRNA) viruses and mediated antigenic adaptive immunity. To understand how TLR7 distinguish pathogen-derived molecular patterns from the host self, it is essential to be able to identify TLR7 receptor interaction interfaces, such as active sites or R848-agonist binding sites. The functional interfaces of TLR7 can serve as targets for structure-based drug design in studying the TLR7 receptor’s structure-function relationship. In contrast to mammalian TLR7, chicken TLR7 (chTLR7) is unknown for its important biological function. Therefore, it has been targeted to mediate contrasting evolutionary patterns of positive selection into non-synonymous SNPs across eleven species using TLR7 conservation patterns (evolutionary conserved and class-specific trace residues), where protein sequence differences to the TLR7 receptors of interest record mutation that have passed positive section across the species. In this study, we characterized the Lys609 residue on chTLR7-ECD homodimer interfaces to reflect the current tendency of evolving positive selection to be transfer into a stabilization direction of the R848-agonist/ chTLR7-ECDs complex under the phylogenetically variable position across species and we suggest a potential indicator for contrasting evolutionary patterns of both the species TLR-ECDs.

Homology Modeling Study of Bovine μ-Calpain Inhibitor-Binding Domains

Chai, Han-Ha,Lim, Dajeong,Lee, Seung-Hwan,Chai, Hee-Yeoul,Jung, Eunkyoung Molecular Diversity Preservation International (MD 2014 INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES Vol.15 No.5

<P>The activated mammalian CAPN-structures, the CAPN/CAST complex in particular, have become an invaluable target model using the structure-based virtual screening of drug candidates from the discovery phase to development for over-activated CAPN linked to several diseases, such as post-ischemic injury and cataract formation. The effect of Ca<SUP>2+</SUP>-binding to the enzyme is thought to include activation, as well as the dissociation, aggregation, and autolysis of small regular subunits. Unfortunately, the Ca<SUP>2+</SUP>-activated enzyme tends to aggregate when provided as a divalent ion at the high-concentration required for the protease crystallization. This is also makes it very difficult to crystallize the whole-length enzyme itself, as well as the enzyme-inhibitor complex. Several parameters that influence CAPN activity have been investigated to determine its roles in Ca<SUP>2+</SUP>-modulation, autoproteolysis, phosphorylation, and intracellular distribution and inhibition by its endogenous inhibitor CAST. CAST binds and inhibits CAPN via its CAPN-inhibitor domains (four repeating domains 1–4; CAST1–4) when CAPN is activated by Ca<SUP>2+</SUP>-binding. An important key to understanding CAPN1 inhibition by CAST is to determine how CAST interacts at the molecular level with CAPN1 to inhibit its protease activity. In this study, a 3D structure model of a CAPN1 bound bovine CAST4 complex was built by comparative modeling based on the only known template structure of a rat CAPN2/CAST4 complex. The complex model suggests certain residues of bovine CAST4, notably, the TIPPKYQ motif sequence, and the structural elements of these residues, which are important for CAPN1 inhibition. In particular, as CAST4 docks near the flexible active site of CAPN1, conformational changes at the interaction site after binding could be directly related to CAST4 inhibitory activity. These functional interfaces can serve as a guide to the site-mutagenesis in research on bovine CAPN1 structure-function relationships for the design of small molecules inhibitors to prevent uncontrolled and unspecific degradation in the proteolysis of key protease substrates.</P>

Molecular Modeling of Small Molecules as BVDV RNA-Dependent RNA Polymerase Allosteric Inhibitors

Chai, Han-Ha,Lim, Dajeong,Chai, Hee-Yeoul,Jung, Eunkyoung Korean Chemical Society 2013 Bulletin of the Korean Chemical Society Vol.34 No.3

Bovine viral diarrhea virus (BVDV), a major pathogen of cattle, is a well-characterized pestivirus which has been used as a good model virus for HCV. The RNA-dependent RNA polymerase (RdRp) plays a key role in the RNA replication process, thus it has been targeted for antivirus drugs. We employed two-dimensional quantitative structure-activity relationship (2D-QSAR) and molecular field analysis (MFA) to identify the molecular substructure requirements, and the particular characteristics resulted in increased inhibitory activity for the known series of compounds to act as effective BVDV inhibitors. The 2D-QSAR study provided the rationale concept for changes in the structure to have more potent analogs focused on the class of arylazoenamines, benzimidazoles, and acridine derivatives with an optimal subset of descriptors, which have significantly contributed to overall anti-BVDV activity. MFA represented the molecular patterns responsible for the actions of antiviral compound at their receptors. We conclude that the polarity and the polarizability of a molecule play a main role in the inhibitory activity of BVDV inhibitors in the QSAR modeling.

T-세포 항원 수용체 매개 신호전달 조절자로서 돼지 CD45RO 구조특성

채한화(Han-Ha Chai),임다정(Dajeong Lim) 한국산학기술학회 2019 한국산학기술학회논문지 Vol.20 No.9

백혈구 공통 항원인 돼지 CD45는 PTPRC 유전자에 암호화 되어 있으며, CD45엑손의 선택적 스플라이싱에 따라 다른 T-세포에서 발현되는 티로신 인산분해효소이다. CD45는 기질인 TCR의 CD3ζ 사슬, Lck, Fyn, Zap-70 kinase의 인산화된 티로신에서 인산을 분해하여 T-세포 항원 수용체(TCR) 매개 신호전달을 조절한다. CD45의 조절이상은 많은 면역 질환과 관련이 있어서, CD45는 면역약물 개발에 표적이 되어왔다. TCR 신호전달의 조절효과를 가진 주요 구조적 특징을 특성화하기 위해, 사람의 알려진 CD45 구조를 템플릿으로 적용하여 돼지 CD45RO(가장 작은 CD45 isoform)의 단백질 구조와 예측된 돼지 CD45RO 모델구조에 CD3ζ 사슬의 ITAM(REEpYDV)를 도킹하여 CD45RO/ITAM 펩타이드 결합구조를 예측하였다. 돼지 CD45RO의 구조적 특징은 세포외영역의 구조견고성과 세포질내 티로신 인산분해효소 도메인의 KNRY와 PTP signature 기능모티프(두 기능 모티프는 ITAM 펩타이드 결합부위의 좁은 입구로 역할)에 있었다. 주요 구조특성은 돼지 CD45RO-ITAM 펩타이드 결합구조 안정성과 결합친화력을 조절하면서 기질 선택성에 영향을 준다. 돼지 CD45RO의 구조적 특성은 T-세포에 특이적인 면역 조절제를 탐색하는 데에 적용될 것이다. Pig CD45, the leukocyte common antigen, is encoded by the PTPRC gene and CD45 is a T cell-type specific tyrosine phosphatase with alternative splicing of its exons. The CD45 is a coordinated regulator of T cell antigen receptor (TCR) signal transduction achieved by dephosphorylating the phosphotyrosine of its substances, including CD3ζ chain of TCR, Lck, Fyn, and Zap-70 kinase. A dysregulation of CD45 is associated with a multitude of immune disease and has been a target for immuno-drug discovery. To characterize its key structural features with the effects of regulating TCR signaling, this study predicted the unknown structure of pig CD45RO (the smallest isoform) and the complex structure bound to the ITAM (REEpYDV) of CD3ζ chain via homology modeling and docking the peptide, based on the known human CD45 structures. These features were integrated into the structural plasticity of extracellular domains and functional KNRY and PTP signature motifs (the role of a narrow entrance into ITAM binding site) of the tyrosine phosphatase domains in a cytoplasmic region from pig CD45RO. This contributes to the selective recognition of phosphotyrosine from its substrates by adjusting the structural stability and binding affinity of the complex. The characterized features of pigCD45RO can be applied in virtual screening of the T-cell specific immunomodulator.

다양한 종에서 하우스키핑 유전자 선택의 중요성

채한화(Han-Ha Chai),노윤정(Yun Jeong Noh),노희종(Hee-Jong Roh),임다정(Dajeong Lim) 한국산학기술학회 2020 한국산학기술학회논문지 Vol.21 No.8

하우스키핑 유전자는 에너지생성, 물질합성, 세포사멸 및 세포방어 등과 같은 세포의 기본적인 기능을 수행하기 때문에 모든 유기체의 세포에서 발현된다. 세포의 기본적인 기능을 유지하기 때문에 발현 수준이 상대적으로 일정하여 단백질 발현 및 목적 유전자의 mRNA 발현 분석 등과 같은 유전자 발현 연구에서 기준 유전자로 사용되고 있다. 그러나 이들 유전자의 발현 수준은 조직과 세포마다 다를 수 있으며, 특정 환경 하에서 변할 수 있다. 그러므로 하우스키핑 유전자의 발현 안정성을 탐색하여 유전자 발현 연구에서 최적의 기준 유전자를 선택하는 것이 중요하다. 이 리뷰는 문헌을 통해 인간, 닭, 돼지 그리고 쥐에서 발견된 하우스키핑 유전자를 요약하고, geNorm, NormFinder 그리고 BestKeeper 소프트웨어를 통해 발현 안정성을 추정하였다. 하우스키핑 유전자의 발현 안정성에 대한 탐색은 유전자 발현 연구에서 실험 조건에 따라 가장 적합한 기준 유전자를 선별할 수 있고, 데이터의 정규화를 위해 적용될 수 있다. Housekeeping genes are expressed in cells of all organisms and perform basic cellular functions such as energy generation, substance synthesis, cell death, and cell defense. Accordingly, the expression levels of housekeeping genes are relatively constant, and thus they are used as reference genes in gene expression studies, such as protein expression and mRNA expression analysis of target genes. However, the levels of expression of these genes may be different among various tissues or cells and may change under certain circumstances. Therefore, it is important to select the best reference gene for specific gene expression research by exploring the stability of housekeeping gene expression. This review summarizes housekeeping genes found in humans, chickens, pigs, and rats in the literature and estimates expression stability using geNorm, NormFinder, and BestKeeper software. The most suitable reference housekeeping gene can selected based on expression stability according to the experimental conditions of the gene expression study and can thus be applied to data normalization.

긴꼬리닭 암컷에서 환우 연관 유전자군 발현 연구

채한화(Han-Ha Chai),노윤정(Yunjeong Noh),최현지(Hyunji Choi),임다정(Dajeong Lim) 한국산학기술학회 2022 한국산학기술학회논문지 Vol.23 No.1

베이지안 모델 기반 약물유사 화합물의 간 독성 예측

채한화(Han-Ha Chai),박원철(Wonchoul Park),진영배(Yeoung Bae Jin),임다정(Dajeong Lim) 한국산학기술학회 2024 한국산학기술학회논문지 Vol.25 No.1

Predicting hepatotoxicity is an important component of safety-related evaluations of drug-like compounds. Hepatotoxicity is related to the physicochemical properties of drug-like compounds, especially their structural alerts. In this study, we developed a Bayesian model to predict the hepatotoxicities of 498 drug-like compounds based on their quantitative structure-toxicity relationships (QSAR). The devised model predicted the hepatotoxicity of these compounds using 25 structural descriptors (such as the ECFP6 fingerprint) and provided a sensitivity, specificity, and concordance of 97.2%, 86.9%, and 90.6%, respectively. The model also successfully classified the 498 drug-like compounds by hepatotoxicity. In addition, TOPKAT<SUP>@</SUP> toxicity models were used to predict hepatotoxic effects related to toxicological endpoints (i.e., LD50, LOAEL) and liver injury-related potentials, such as Ames mutagenicity, carcinogenicity in male and female mice, and developmental toxicity potentials. Our results indicate that combined use of the devised Bayesian hepatotoxicity prediction model and the TOPKAT<SUP>@</SUP> hepatotoxicity model could replace experimental safety assessments of drug-like compounds. Accordingly, adopting the devised Bayesian toxicity model would reduce the time and cost of animal-based toxicity research by considering the safety and effectiveness of candidate compounds.