• Bulletin of the Korean Chemical Society
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• 제34권5호
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• pp.1401-1406
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• 2013

This study tested the feasibility of observing H/D exchange of intact protein by top-down electron capture dissociation (ECD) mass spectrometry for the investigation of protein structure. Ubiquitin is selected as a model system. Local structural information was obtained from the deuteration levels of c and $z^{\cdot}$ ions generated from ECD. Our results showed that ${\alpha}$-helix region has the lowest deuteration level and the C-terminal fraction containing a highly mobile tail has the highest deuteration level, which correlates well with previous X-Ray and HDX/NMR analyses. We studied site-specific H/D exchange kinetics by monitoring H/D exchange rate of several structural motives of ubiquitin. Two hydrogen bonded ${\beta}$-strands showed similar HDX rates. However, the outer ${\beta}$-strand always has higher deuteration level than the inner ${\beta}$-strand. The HDX rate of the turn structure (residues 8-11) is lower than that of ${\beta}$-strands (residues 1-7 and residues 12-17) it connects. Although isotopic distribution gets broader after H/D exchange which results in a limited number of backbone cleavage sites detected, our results demonstrate that this method can provide valuable detailed structural information of proteins. This approach should also be suitable for the structural investigation of other unknown proteins, protein conformational changes, as well as protein-protein interactions and dynamics.

• 한국자기공명학회논문지
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• 제19권3호
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• pp.112-118
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• 2015

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 $60^{\circ}C$ 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.

• Journal of Photoscience
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• 제11권3호
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• pp.95-99
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• 2004

Optical absorption and emission studies have been carried out to understand the effect of deuterium on the solvent dependent photophysical properties of curcumin in deuterated solvents such as $CDCl_3,\;(CD_3)_2SO,\;(CD_3)_2CO,\;CD_3OD\;and\;CD_3CN$. Optical absorption spectral studies showed that there is no significant shift in absorption maxima compared to the non-deuterated solvent. The fluorescence maxima shows significant shift with polarity of solvent but not much affected by the deuteration. The fluorescence quantum yield of curcumin increased marginally in almost all the deuterated solvents, indicating reduction in the non-radiative pathways. The fluorescence decay was biexponential in all the solvents and the average fluorescence lifetime was not much affected with deuteration, but showed decrease with increasing solvent polarity. Based on these studies, it is concluded that intermolecular hydrogen transfer is only partially responsible for the excited state deactivation of curcumin.

• Bulletin of the Korean Chemical Society
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• 제23권5호
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• pp.727-735
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• 2002

Proton coupled carbon-13 relaxation experiment was performed to investigate the effect of vicinal protons on spin-lattice relaxation of methylene carbon-13 in n-undecane. A BIRD type pulse sequence was employed as a way to check the validity of describing the 13CH2 moiety as an isolated AX2 spin system. The results show that the presence of vicinal protons exerts substantial influence on the relaxation of methylene carbon-13, indicating that it is not a very good approximation to treat a methylene moiety as an isolated AX2 spin system.

• 분석과학
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• 제7권2호
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• pp.213-224
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• 1994

선택적으로 중수소를 치환시킨 싸이클로옥타논의 여러 동위원소 이성질체들을 합성하였다. 고유동위원소 효과에 의해 영향을 받는 $^{13}C$ NMR 화학적 이동값들을 각 이성질체에 대해 저온에서 계통적으로 관찰하였다. 특히 싸이클로옥타논이 선호하는 안정한 형태 이성질체인 클 boat-chair 형과 연관시켜 이 효과들을 논의하였다.

• 대한화학회지
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• 제28권1호
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• pp.14-19
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• 1984

메트미오글로빈의 압력에 의한 변성의 $H_2O$와 $D_2O$에서의 차이를 pH 5.7과 pH 7.0에서 연구하였다. 메트미오글로빈은 $D_2O$에서 $H_2O$보다 더 작은 압력에서 변성하였다. 그 차이가 pH 5.7에서가 pH 7에서 보다 더 컸다. $H_2O$와 $D_2O$에서 이 안정도의 차이가 단백질에 대한 $H^+$와 $D^+$의 결합에 차이가 있기 때문이며 또한, 수소가 중수소로 바뀜에 따르는 구조 변화 때문인 것으로 사료된다.

• 대한화학회지
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• 제33권2호
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• pp.238-246
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• 1989

옥사티올란 고리에 대해서 술폭시드기와 2-메틸기가 동일 평면상에 놓인 1,3-옥사티올란술폭시드 4는 sigmatropic 전위를 통하여 고리확대 생성물로 전환한다. 이 생성물의 구조는 디히드로-1,4-옥사티인 6 또는 이것의 구조 이성체인 exo형 화합물 7의 가능성이 있다. 본 연구에서는 물리적 및 화학적 방법에 의하여 두 alternative 구조 중 올바른 구조를 밝혔다. 즉 수소핵자기 공명스펙트럼, 자외선 흡수스펙트럼, 그리고 질량분석 스펙트럼으로부터 실제로 얻어진 화합물은 디히드로옥시티인 6이었다. 또한 중수소 치환반응으로부터 환팽창 반응에서 처음 exo형 화합물 7이 생성되나 tautomerization에 의해서 결국 디히드로-1,4-옥사티인 6으로 전환함을 알았다.

• Journal of Photoscience
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• 제1권1호
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• pp.9-14
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• 1994

Exposure of isolated intact mitochondria to near UV to visible light resulted in not only loss of respiration, the most well-documented phenomenon regarding phototoxic effects in the respiring organelles, but also lipid peroxidation of membranes and mitochondrial swelling; these turned out to be O$_2$-dependent and thus prevented by anaerobiosis, enhanced by a partial deuteration of the suspension medium, and suppressed by the presence of a singlet oxygen ($^1O_2$) scavenger. Measurements of the spectral dependence of such detrimental effects of light on mitochondrial structure and function revealed that all the resulting spectra bear a significant resemblance to the action spectrum for photogeneration of $^1O_2$ from mitochondrial membranes, which in turn carries the spectral characteristics of light absorption by mitochondrial Fe-S centers. Futhermore, destructing the Fe-S centers by a mercurial treatment of mitochondria brought about a striking reduction of the light-induced membrane peroxidation and swelling of mitochondria. These results are consistent with the suggestion that the impairment of functional, structural integrity of mitochondria caused by strong irradiation is directly related to the production of $^1O_2$ in mitochondria, photosensitized by the Fe-S centers. This paper also presents kinetic data which indicate that, among various membrane-bound protein systems associated with mitochondrial energy metabolism, the respiratory chain is the primary target for photodamage.

• 한국자기공명학회논문지
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• 제19권3호
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• pp.137-142
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• 2015

NMR-based structural studies on membrane proteins are appreciated quite challenging due to various reasons, generally including the narrow dispersion of NMR spectra, the severe peak broadening, and the lack of long range NOEs. In spite of the poor biophysical properties, structural studies on membrane proteins have got to go on, considering their functional importance in biological systems. In this review, we provide a simple overview of the techniques generally used in structural studies of membrane proteins by solution NMR, with experimental examples of a helical membrane protein, caveolin 3. Detergent screening is usually employed as the first step and the selection of appropriate detergent is the most important for successful approach to membrane proteins. Various tools can then be applied as specialized NMR techniques in solution that include sample deteuration, amino-acid selective isotope labeling, residual dipolar coupling, and paramagnetic relaxation enhancement.

• 천문학논총
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• 제29권2호
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• pp.29-34
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• 2014

We have observed the deuterated methanol, $CH_3OD$, toward the hot core MM1 in the massive star-forming region DR21 (OH) using the Submillimeter Array with a high angular resolution of about 1 arcsecond. The position of the hot core associated with the sub-core MM1a was confirmed to coincide with the continuum peak where an embedded young stellar object is located. The column density of $CH_3OD$ was found to be about $(2{\pm}1){\times}10^{16}cm^{-2}$ toward the MM1a center. The abundance ratio $CH_3OD/CH_3OH$ was measured to be ~ 0.45, which is about the median value for low mass star-forming cores but much larger than those of the massive star-forming cores. The ratio is believed to change depending on, for example, the chemical condition, the temperature and the density of the source. This ratio may further depend on the evolutionary phase especially in the massive-star-forming cores. The sub-core MM1a is thought to be in the very early phase of star formation. This large abundance ratio found in this source indicates that even the massive star-forming cores, during a relatively short period in the very early stage of star formation, may also show a chemical state resulted from the cold and dense pre-collapsing phase, the enhanced deuteration as found in low mass star-forming cores.