Pub Date : 2016-03-20DOI: 10.6564/JKMRS.2016.20.1.001
S. Park
The proton NMR line widths and spin-lattice relaxation rates, T1, of ferroelectric LiH3(SeO3)2, Li2SO4·H2O, and LiN2H5SO4 single crystals were measured as a function of temperature. The line width measurements reveal rigid lattice behavior of all the crystals at low temperatures and line narrowing due to molecular motion at higher temperatures. The temperature dependences of the proton T1 for these crystals exhibit maxima, which are attributed to the effects of molecular motion by the Bloembergen Purcell Pound theory. The activation energies for the molecular motions of H in these crystals were obtained. From these analysis, H in LiH3(SeO3)2 undergoes molecular motion more easily than H in LiN2H5SO4 and Li2SO4·H2O crystals.
{"title":"Study of molecular motion by 1 H NMR relaxation in ferroelectric LiH 3 (SeO 3 ) 2 , Li 2 SO 4 ·H 2 O, and LiN 2 H 5 SO 4 single crystals","authors":"S. Park","doi":"10.6564/JKMRS.2016.20.1.001","DOIUrl":"https://doi.org/10.6564/JKMRS.2016.20.1.001","url":null,"abstract":"The proton NMR line widths and spin-lattice relaxation rates, T1, of ferroelectric LiH3(SeO3)2, Li2SO4·H2O, and LiN2H5SO4 single crystals were measured as a function of temperature. The line width measurements reveal rigid lattice behavior of all the crystals at low temperatures and line narrowing due to molecular motion at higher temperatures. The temperature dependences of the proton T1 for these crystals exhibit maxima, which are attributed to the effects of molecular motion by the Bloembergen Purcell Pound theory. The activation energies for the molecular motions of H in these crystals were obtained. From these analysis, H in LiH3(SeO3)2 undergoes molecular motion more easily than H in LiN2H5SO4 and Li2SO4·H2O crystals.","PeriodicalId":17414,"journal":{"name":"Journal of the Korean magnetic resonance society","volume":"43 1","pages":"1-6"},"PeriodicalIF":0.3,"publicationDate":"2016-03-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"71329919","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2016-03-20DOI: 10.6564/JKMRS.2016.20.1.007
Gilhoon Kim, Hoshik Won
glycol) is a polymer compound widely used as a wide range of applications in the textile industry. PTMEG sub-stance carrying various 1,800~2,000 molecular weight are mainly used as the raw material of the spandex production. Molecular weight and degree of polymerization value for 4 different PTMEG samples under pilot plant scale synthetic process were determined by a new quantitative NMR method. In NMR experiments, p -toluenesulfonic acid(TSOH) was used for external standard material of PTMEG quantitative analysis. were measuring The concentration of the primary standard TSOH was measured by UV/Vis spectroscopy. By using NMR peak assignments and the integral values of designated proton NMR peaks, We were able to measure the % composition of the synthetic PTMEG polymers, concentrations, molecular weight and the degree of polymerization that show the synthetic process of each manufacturing pilot plant. By utilizing a newly developed quantitative NMR method were able to obtain the molecular weight of PTMEG samples within 0.08 error % range.
{"title":"Quantitative NMR Analysis of PTMEG compounds","authors":"Gilhoon Kim, Hoshik Won","doi":"10.6564/JKMRS.2016.20.1.007","DOIUrl":"https://doi.org/10.6564/JKMRS.2016.20.1.007","url":null,"abstract":"glycol) is a polymer compound widely used as a wide range of applications in the textile industry. PTMEG sub-stance carrying various 1,800~2,000 molecular weight are mainly used as the raw material of the spandex production. Molecular weight and degree of polymerization value for 4 different PTMEG samples under pilot plant scale synthetic process were determined by a new quantitative NMR method. In NMR experiments, p -toluenesulfonic acid(TSOH) was used for external standard material of PTMEG quantitative analysis. were measuring The concentration of the primary standard TSOH was measured by UV/Vis spectroscopy. By using NMR peak assignments and the integral values of designated proton NMR peaks, We were able to measure the % composition of the synthetic PTMEG polymers, concentrations, molecular weight and the degree of polymerization that show the synthetic process of each manufacturing pilot plant. By utilizing a newly developed quantitative NMR method were able to obtain the molecular weight of PTMEG samples within 0.08 error % range.","PeriodicalId":17414,"journal":{"name":"Journal of the Korean magnetic resonance society","volume":"20 1","pages":"7-12"},"PeriodicalIF":0.3,"publicationDate":"2016-03-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"71329512","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2015-12-20DOI: 10.6564/JKMRS.2015.19.3.143
Won-Je Kim, Nak-Kyoon Kim
Abstract In some pathogenic bacteria, there are RNA thermometers, which regulate the production of virulence associated factors or heat shock proteins depending on temperature changes. Like a riboswitches, RNA thermometers are located in the 5’-untranslated region and involved translational gene regulatory mechanism. RNA thermometers block the ribosome-binding site and start codon area under the 37℃ living systems. within their secondary structure. After bacterial infection, increased the temperature in the host causes conformations changes of RNA, and the ribosome-binding site is exposed for translational initiation. Because structural differences between open and closed forms of RNA thermometers are mainly mediated by base pairing changes, NMR spectroscopy is a very useful method to study these thermodynamically changing RNA structure. In this review, we briefly provide a fundamental function of RNA thermometers, and also suggest a proper NMR experiments for studying RNA thermometers.
{"title":"Proper NMR methods for studying RNA thermometers","authors":"Won-Je Kim, Nak-Kyoon Kim","doi":"10.6564/JKMRS.2015.19.3.143","DOIUrl":"https://doi.org/10.6564/JKMRS.2015.19.3.143","url":null,"abstract":"Abstract In some pathogenic bacteria, there are RNA thermometers, which regulate the production of virulence associated factors or heat shock proteins depending on temperature changes. Like a riboswitches, RNA thermometers are located in the 5’-untranslated region and involved translational gene regulatory mechanism. RNA thermometers block the ribosome-binding site and start codon area under the 37℃ living systems. within their secondary structure. After bacterial infection, increased the temperature in the host causes conformations changes of RNA, and the ribosome-binding site is exposed for translational initiation. Because structural differences between open and closed forms of RNA thermometers are mainly mediated by base pairing changes, NMR spectroscopy is a very useful method to study these thermodynamically changing RNA structure. In this review, we briefly provide a fundamental function of RNA thermometers, and also suggest a proper NMR experiments for studying RNA thermometers.","PeriodicalId":17414,"journal":{"name":"Journal of the Korean magnetic resonance society","volume":"19 1","pages":"143-148"},"PeriodicalIF":0.3,"publicationDate":"2015-12-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"71329903","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2015-12-20DOI: 10.6564/JKMRS.2015.19.3.149
Ja-shil Hyun, Sung Jean Park
Abstract Acyl carrier protein is related with fatty acid biosynthesis in whichspecific enzymes are in-volved. Especially, acyl carrier protein (ACP) is the key component in the growing of fatty acid chain. ACP is the small, very acidic protein that covalently binds various intermediates of fatty acyl chain. Acyl-ation of ACP is mediated by holoacyl carrier protein -synthase (ACPS), which transfers the 4’PP-moiety of CoA to the 36th residue Ser of apoACP. Acyl carrier protein P (ACPP) is one of ACPs from Helicobacter plyori. The NMR structure of ACPP consists of four helices, which were reported previously. Here we show how acylation of ACPP can affect the overall structure of ACPP and figured out the contact surface of ACPP to acyl chain attached during expression of ACPP in E. coli Based on the chemical shift . r- pertubation data, the acylation of ACCP seems to affect the conformation of the long loop connecting helix I and helix II as well as the second short loop connect-ing helix II and helix III. The significant chemical shift change of Ile 54 upon acylation supports the contact of acyl chain and the second loop.
{"title":"Effect of Acylation on the Structure of the Acyl Carrier Protein P","authors":"Ja-shil Hyun, Sung Jean Park","doi":"10.6564/JKMRS.2015.19.3.149","DOIUrl":"https://doi.org/10.6564/JKMRS.2015.19.3.149","url":null,"abstract":"Abstract Acyl carrier protein is related with fatty acid biosynthesis in whichspecific enzymes are in-volved. Especially, acyl carrier protein (ACP) is the key component in the growing of fatty acid chain. ACP is the small, very acidic protein that covalently binds various intermediates of fatty acyl chain. Acyl-ation of ACP is mediated by holoacyl carrier protein -synthase (ACPS), which transfers the 4’PP-moiety of CoA to the 36th residue Ser of apoACP. Acyl carrier protein P (ACPP) is one of ACPs from Helicobacter plyori. The NMR structure of ACPP consists of four helices, which were reported previously. Here we show how acylation of ACPP can affect the overall structure of ACPP and figured out the contact surface of ACPP to acyl chain attached during expression of ACPP in E. coli Based on the chemical shift . r- pertubation data, the acylation of ACCP seems to affect the conformation of the long loop connecting helix I and helix II as well as the second short loop connect-ing helix II and helix III. The significant chemical shift change of Ile 54 upon acylation supports the contact of acyl chain and the second loop.","PeriodicalId":17414,"journal":{"name":"Journal of the Korean magnetic resonance society","volume":"19 1","pages":"149-155"},"PeriodicalIF":0.3,"publicationDate":"2015-12-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"71329911","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2015-12-20DOI: 10.6564/JKMRS.2015.19.3.099
Yoo-Sup Lee, Ji-hoon Kim, M. Seo, K. Ryu, Eun-hee Kim, H. Won
Hsp33 is a prokaryotic molecular chaperon that exerts a holdase activity upon response to an oxidative stress at raised temperature. In particular, intramolecular disulfide bond formation between the four conserved cysteines that bind a zinc ion in reduced state is known to be critically associated with the redox sensing. Here we report the backbone NMR assignment results of the half-oxidized Hsp33, where only two of the four cysteines form an intramolecular disulfide bond. Almost all of the resolved peaks could be unambiguously assigned, although the total assignments extent reached just about 50%. Majority of the missing assignments could be attributed to a significant spectral collapse, largely due to the oxidation-induced unfolding of the C-terminal redox-switch domain. These results support two previous suggestions: conformational change in the first oxidation step is localized mainly in the C-terminal zinc-binding domain, and the half-oxidized form would be still inactive. However, some additional regions appeared to be potentially changed from the reduced state, which suggest that the half-oxidized conformation would be an intermediate state that is more labile to heat and/or further oxidation.
{"title":"Oxidation-induced conformational change of Hsp33, monitored by NMR","authors":"Yoo-Sup Lee, Ji-hoon Kim, M. Seo, K. Ryu, Eun-hee Kim, H. Won","doi":"10.6564/JKMRS.2015.19.3.099","DOIUrl":"https://doi.org/10.6564/JKMRS.2015.19.3.099","url":null,"abstract":"Hsp33 is a prokaryotic molecular chaperon that exerts a holdase activity upon response to an oxidative stress at raised temperature. In particular, intramolecular disulfide bond formation between the four conserved cysteines that bind a zinc ion in reduced state is known to be critically associated with the redox sensing. Here we report the backbone NMR assignment results of the half-oxidized Hsp33, where only two of the four cysteines form an intramolecular disulfide bond. Almost all of the resolved peaks could be unambiguously assigned, although the total assignments extent reached just about 50%. Majority of the missing assignments could be attributed to a significant spectral collapse, largely due to the oxidation-induced unfolding of the C-terminal redox-switch domain. These results support two previous suggestions: conformational change in the first oxidation step is localized mainly in the C-terminal zinc-binding domain, and the half-oxidized form would be still inactive. However, some additional regions appeared to be potentially changed from the reduced state, which suggest that the half-oxidized conformation would be an intermediate state that is more labile to heat and/or further oxidation.","PeriodicalId":17414,"journal":{"name":"Journal of the Korean magnetic resonance society","volume":"19 1","pages":"99-105"},"PeriodicalIF":0.3,"publicationDate":"2015-12-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"71329730","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2015-12-20DOI: 10.6564/JKMRS.2015.19.3.119
H. Yoon, A. Lim
The thermodynamic properties and structural geometry of KMgCl 3 ·6H 2 O were investigated using thermogravimetric analysis, differential scanning calorimetry, and nuclear magnetic resonance. The initial mass loss occurs around 351 K (=T d ), which is interpreted as the onset of partial thermal decomposition. Phase transition temperatures were found at 435 K (=T C1 ) and 481 K (=T C2 ). The temperature dependences of the spin–lattice relaxation time T 1 for the 1 H nucleus changes abruptly near T C1 . These changes are associated with changes in the geometry of the arrangement of octahedral water molecules.
采用热重分析、差示扫描量热法和核磁共振等方法研究了kmgcl3·6h2o的热力学性质和结构几何形状。初始质量损失发生在351 K (=T d)左右，这被解释为部分热分解的开始。相变温度分别为435 K (=T C1)和481 K (=T C2)。氢原子核的自旋-晶格弛豫时间t1的温度依赖性在t1附近突然改变。这些变化与八面体水分子排列的几何变化有关。
{"title":"Thermodynamic properties and structural geometry of KMgCl 3 ·6H 2 O single crystals","authors":"H. Yoon, A. Lim","doi":"10.6564/JKMRS.2015.19.3.119","DOIUrl":"https://doi.org/10.6564/JKMRS.2015.19.3.119","url":null,"abstract":"The thermodynamic properties and structural geometry of KMgCl 3 ·6H 2 O were investigated using thermogravimetric analysis, differential scanning calorimetry, and nuclear magnetic resonance. The initial mass loss occurs around 351 K (=T d ), which is interpreted as the onset of partial thermal decomposition. Phase transition temperatures were found at 435 K (=T C1 ) and 481 K (=T C2 ). The temperature dependences of the spin–lattice relaxation time T 1 for the 1 H nucleus changes abruptly near T C1 . These changes are associated with changes in the geometry of the arrangement of octahedral water molecules.","PeriodicalId":17414,"journal":{"name":"Journal of the Korean magnetic resonance society","volume":"19 1","pages":"119-123"},"PeriodicalIF":0.3,"publicationDate":"2015-12-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"71329963","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2015-12-20DOI: 10.6564/JKMRS.2015.19.3.124
Hyeji Ko, Gyeonghyeon Gong, Gijin Jeong, Ikjang Choi, Hyeonglim Seo, Youngbok Lee
Hyperpolarization methods are the most emerging techniques in the field of magnetic resonance (MR) researches since they make a contribution to overcoming sensitivity limitation of MR spectroscopy and imaging, leading to new fields of researches, real-time in vivo metabolic/molecular imaging and MR analysis of chemical/biological reactions in non-equilibrium conditions. Make use of enormous signal enrichments, it becomes feasible to investigate various chemical and biochemical systems with low γ nuclei in real-time. This review deals with the theoretical principals of common hyperpolarization methods and their experimental features. In addition, more detailed theories, mechanisms, and applications of dissolution dynamic nuclear polarization (D-DNP) are discussed.
{"title":"Hyperpolarization: Sensitivity Boost in Magnetic Resonance Spectroscopy and Imaging","authors":"Hyeji Ko, Gyeonghyeon Gong, Gijin Jeong, Ikjang Choi, Hyeonglim Seo, Youngbok Lee","doi":"10.6564/JKMRS.2015.19.3.124","DOIUrl":"https://doi.org/10.6564/JKMRS.2015.19.3.124","url":null,"abstract":"Hyperpolarization methods are the most emerging techniques in the field of magnetic resonance (MR) researches since they make a contribution to overcoming sensitivity limitation of MR spectroscopy and imaging, leading to new fields of researches, real-time in vivo metabolic/molecular imaging and MR analysis of chemical/biological reactions in non-equilibrium conditions. Make use of enormous signal enrichments, it becomes feasible to investigate various chemical and biochemical systems with low γ nuclei in real-time. This review deals with the theoretical principals of common hyperpolarization methods and their experimental features. In addition, more detailed theories, mechanisms, and applications of dissolution dynamic nuclear polarization (D-DNP) are discussed.","PeriodicalId":17414,"journal":{"name":"Journal of the Korean magnetic resonance society","volume":"19 1","pages":"124-131"},"PeriodicalIF":0.3,"publicationDate":"2015-12-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"71329969","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2015-12-20DOI: 10.6564/JKMRS.2015.19.3.106
Kiyoung Lee, Ji-Hun Kim, Ye-Ji Bae, Bong‐Jin Lee
Abstract Helicobacter pylori H. pylori ( ) survives in acidic and fluctuating pH conditions of the stomach. The pH effect on H. pylori proteins is important for the advanced understanding of its evolution and viability, although this bacterium has the molecular machinery that neutralizes theacidic condition. HP1492 is known as a conserved NifU–like protein from H. pylori. NifU is a nitrogen fixation protein that mediates the transfer of iron–sulfur (Fe–S) cluster to iron–sulfur proteins like ferredoxin. Commonly, the monomeric reduced state of NifU can be converted to the dimeric oxidized state by intermolecular disulfide bond formation. Because it remains unclear that HP1492 actually behaves as known NifU protein, we first found that this protein can adopt both oxidized and reduced forms using size exclusion chromatography. Circular dichroism experiment showed that HP1492 is relatively well–structured at pH 6.5, compared to other pH conditions. On the basis of the backbone resonance assignment of HP1492, we further characterized the residues that are sensitive to pH using NMR spectroscopy. These residues showing large chemical shift changes could be mapped onto the secondary structure of the protein. Our results could provide the foundation for structural and biophysical studies on a wide spectrum of NifU proteins.
{"title":"pH Effect on the Structure of Reduced NifU-like Protein from Helicobacter pylori","authors":"Kiyoung Lee, Ji-Hun Kim, Ye-Ji Bae, Bong‐Jin Lee","doi":"10.6564/JKMRS.2015.19.3.106","DOIUrl":"https://doi.org/10.6564/JKMRS.2015.19.3.106","url":null,"abstract":"Abstract Helicobacter pylori H. pylori ( ) survives in acidic and fluctuating pH conditions of the stomach. The pH effect on H. pylori proteins is important for the advanced understanding of its evolution and viability, although this bacterium has the molecular machinery that neutralizes theacidic condition. HP1492 is known as a conserved NifU–like protein from H. pylori. NifU is a nitrogen fixation protein that mediates the transfer of iron–sulfur (Fe–S) cluster to iron–sulfur proteins like ferredoxin. Commonly, the monomeric reduced state of NifU can be converted to the dimeric oxidized state by intermolecular disulfide bond formation. Because it remains unclear that HP1492 actually behaves as known NifU protein, we first found that this protein can adopt both oxidized and reduced forms using size exclusion chromatography. Circular dichroism experiment showed that HP1492 is relatively well–structured at pH 6.5, compared to other pH conditions. On the basis of the backbone resonance assignment of HP1492, we further characterized the residues that are sensitive to pH using NMR spectroscopy. These residues showing large chemical shift changes could be mapped onto the secondary structure of the protein. Our results could provide the foundation for structural and biophysical studies on a wide spectrum of NifU proteins.","PeriodicalId":17414,"journal":{"name":"Journal of the Korean magnetic resonance society","volume":"19 1","pages":"106-111"},"PeriodicalIF":0.3,"publicationDate":"2015-12-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"71329788","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2015-12-20DOI: 10.6564/JKMRS.2015.19.3.112
Jihong Kim, Dongwook Choi, Chankyu Park, K. Ryu
Hsp31 protein is one of the members of DJ-1 superfamily proteins and has a dimeric structure of which molecular weight (MW) is 62 kDa. The mutation of DJ-1 is closely related to early onset of Parkinson’s disease. Hsp31 displays Zn +2 -binding activity and was first reported to be a holding chaperone in E. coli . Its additional glyoxalase III active has recently been characterized. Moreover, an incubation at 60 ° C induces Hsp31 protein to form a high MW oligomer (HMW) in vitro , which accomplishes an elevated holding chaperone activity. The NMR technique is elegant method to probe any local or global structural change of a protein in responses to environmental stresses (heat, pH, and metal). Although the presence of the backbone chemical shifts (bbCSs) is a prerequisite for detailed NMR analyses of the structural changes, general HSQC-based triple resonance experiments could not be used for 62 kDa Hsp31 protein. Here, we prepared the per-deuterated Hsp31 and performed the TROSY-based triple resonance experiments for the bbCSs assignment. Here, detailed processes of per-deuteration and the NMR experiments are described for other similar NMR approaches.
{"title":"Per-deuteration and NMR experiments for the backbone assignment of 62 kDa protein, Hsp31","authors":"Jihong Kim, Dongwook Choi, Chankyu Park, K. Ryu","doi":"10.6564/JKMRS.2015.19.3.112","DOIUrl":"https://doi.org/10.6564/JKMRS.2015.19.3.112","url":null,"abstract":"Hsp31 protein is one of the members of DJ-1 superfamily proteins and has a dimeric structure of which molecular weight (MW) is 62 kDa. The mutation of DJ-1 is closely related to early onset of Parkinson’s disease. Hsp31 displays Zn +2 -binding activity and was first reported to be a holding chaperone in E. coli . Its additional glyoxalase III active has recently been characterized. Moreover, an incubation at 60 ° C induces Hsp31 protein to form a high MW oligomer (HMW) in vitro , which accomplishes an elevated holding chaperone activity. The NMR technique is elegant method to probe any local or global structural change of a protein in responses to environmental stresses (heat, pH, and metal). Although the presence of the backbone chemical shifts (bbCSs) is a prerequisite for detailed NMR analyses of the structural changes, general HSQC-based triple resonance experiments could not be used for 62 kDa Hsp31 protein. Here, we prepared the per-deuterated Hsp31 and performed the TROSY-based triple resonance experiments for the bbCSs assignment. Here, detailed processes of per-deuteration and the NMR experiments are described for other similar NMR approaches.","PeriodicalId":17414,"journal":{"name":"Journal of the Korean magnetic resonance society","volume":"51 1","pages":"112-118"},"PeriodicalIF":0.3,"publicationDate":"2015-12-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"71329801","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2015-12-20DOI: 10.6564/JKMRS.2015.19.3.132
Jongsoo Lim
Nuclear magnetic resonance (NMR) spectroscopy, owing to its ability to provide atomic level information on molecular structure, dynamics and interaction, has become one of the most powerful methods in early drug discovery where hit finding and hit-to-lead generation are mainly pursued. In recent years, drug discovery programs originating from the fragment-based drug discovery (FBDD) strategies have been widely incorporated into academia and industry in which a wide variety of NMR methods become an indispensable arsenal to elucidate the binding of small molecules onto bimolecular targets. In this review, I briefly describe FBDD and introduce NMR methods mainly used in FBDD campaigns of my company. In addition, quality control of fragment library and practical NMR methods in industrial aspect are discussed shortly.
{"title":"NMR methods in fragment based drug discovery","authors":"Jongsoo Lim","doi":"10.6564/JKMRS.2015.19.3.132","DOIUrl":"https://doi.org/10.6564/JKMRS.2015.19.3.132","url":null,"abstract":"Nuclear magnetic resonance (NMR) spectroscopy, owing to its ability to provide atomic level information on molecular structure, dynamics and interaction, has become one of the most powerful methods in early drug discovery where hit finding and hit-to-lead generation are mainly pursued. In recent years, drug discovery programs originating from the fragment-based drug discovery (FBDD) strategies have been widely incorporated into academia and industry in which a wide variety of NMR methods become an indispensable arsenal to elucidate the binding of small molecules onto bimolecular targets. In this review, I briefly describe FBDD and introduce NMR methods mainly used in FBDD campaigns of my company. In addition, quality control of fragment library and practical NMR methods in industrial aspect are discussed shortly.","PeriodicalId":17414,"journal":{"name":"Journal of the Korean magnetic resonance society","volume":"19 1","pages":"132-136"},"PeriodicalIF":0.3,"publicationDate":"2015-12-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"71329845","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
京公网安备 11010802042870号
京ICP备2023020795号-1
扫码关注我们
求助内容：
应助结果提醒方式：