• 제목/요약/키워드: pH dependent

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형광 단백질 mCherry-I202T의 pH 감응성 분석 (Characterization of pH Dependent Properties of mCherry Mutant, I202T)

  • 이상민;정민섭
    • 공업화학
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    • 제32권1호
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    • pp.10-14
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    • 2021
  • DsRed에서 유래한 적색 형광 단백질들 중 하나인 mCherry는 GFP와 유사한 3차 구조를 가진 잘 알려진 적색 형광 단백질 중 하나이며, 수소결합 네트워크를 형성하고 있지 않아서 pH 변화에 민감하지 않다. 반면 mCherry의 발색단 근처에서 돌연변이를 야기 시켜 만든 mCherry-I202T는 mCherry 단백질의 202번째 아미노산인 이소루신(Ile)을 트레오닌(Thr)으로 치환함으로써 모체와는 다르게 추가적인 수소결합을 형성하여, 주위 pH에 더욱 민감하게 반응할 뿐 아니라 적색 편이된 형광을 보였다. 수소결합이 확장된 I202T의 pH 민감성을 검증하기 위해 산성과 염기성 pH 범위에서 I202T의 UV-vis 흡광 스펙트럼 변화와 가역성을 확인하고, 그를 pH sensor에 적용 가능한지 그 가능성을 검증하고자 하였다.

In vitro inhibitory activities of urea analogues on bacterial urease

  • Chang, Pan-Sup;Shu, Byung-Se;Nancy A.Strockbine;Kunin, Galvin M
    • Archives of Pharmacal Research
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    • 제9권3호
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    • pp.163-167
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    • 1986
  • Twenty six urea analogues, most of which have already been approved for human use, were tested for their antiurease activity in vitro. Cell-free extracts obtained from a clinical isolate of Proteus mirabilis was used as the source of enzyme. Acetohydroxamic acid which is a proven potent urease inhibitor but not approved for human use was again shown to be the most active compound among the tested. Phenacemide, cycloserine, and deferoxamine were demonstrated to be moderate inhibitors. Oxtetracycline, trimethoprim, and cefamandole revealed a demonstrable antiruease activity, but only at very high concentrations. The antiurease activity of cycloserine, trimethoprim, and cefamandole was pH dependent-only active at acidic pH. The inhibitory activity of acetohydroxamic acid however was independent of change in pH. The inhibitory activity of acetohydroxamic acid however was independent of change in pH. Hydrogen ion concentration plays an important role in urease activity and acidification (pH 5. 5) alone eliminates approximately 65% of the enzymic activity. Adjustment of pH therefore appears to be an important adjunct in reducing unrease activity and should always be studied to maximize the effcacy of antiurease compounds under investigation.

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pH-Dependent surface-enhanced resonance Raman scattering of yeast iso-1-cytochrome c adsorbed on silver nanoparticle surfaces under denaturing conditions at pH < 3

  • Lee, So-Yeong;Joo, Sang-Woo;Lee, Seong-Hoon;Lim, Man-Ho
    • BMB Reports
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    • 제42권4호
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    • pp.223-226
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    • 2009
  • We measured the pH-induced spectral changes of yeast iso-1-cytochrome c on silver nanoparticle surfaces using surface-enhanced resonance Raman scattering (SERRS) at 457.9 nm. At a pH of ~3, the Met80 ligand in yeast iso-1-cytochrome c is assumed to dissociate, leading to a marked conformational change as evidenced by the vibrational spectral shifts. The Soret band at ~410 nm in the UV-Vis spectrum shifted to ~396 nm at pH~3, indicating a transition from a low spin state to a high spin state from a weak interaction with a water molecule. Thus, SERRS spectroscopy can measure the pH-induced denaturalization of cyt c adsorbed on metal nanoparticle surfaces at a lower concentration with a better sensitivity than ordinary resonance Raman spectroscopy.

Thermal Stability of Phaseolus vulgaris Leucoagglutinin: a Differential Scanning Calorimetry Study

  • Biswas, Shyamasri;Kayastha, Arvind M.
    • BMB Reports
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    • 제35권5호
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    • pp.472-475
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    • 2002
  • Phaseolus vulgaris phytohemagglutinin L is a homotetrameric-leucoagglutinating seed lectin. Its three-dimensional structure shows similarity with other members of the legume lectin family. The tetrameric form of this lectin is pH dependent. Gel filtration results showed that the protein exists in its dimeric state at pH 2.5 and as a tetramer at pH 7.2. Contrary to earlier reports on legume lectins that possess canonical dimers, thermal denaturation studies show that the refolding of phytohemagglutinin L at neutral pH is irreversible. Differential scanning calorimetry (DSC) was used to study the denaturation of this lectin as a function of pH that ranged from 2.0 to 3.0. The lectin was found to be extremely thermostable with a transition temperature around $82^{\circ}C$ and above $100^{\circ}C$ at pH 2.5 and 7.2, respectively. The ratio of calorimetric to vant Hoff enthalpy could not be calculated because of its irreversible-folding behavior. However, from the DSC data, it was discovered that the protein remains in its compact-folded state, even at pH 2.3, with the onset of denaturation occurring at $60^{\circ}C$.

Compressibility and hydraulic conductivity of calcium bentonite treated with pH-responsive polymer

  • Choo, Hyunwook;Choi, Youngmin;Kim, Young-Uk;Lee, Woojin;Lee, Changho
    • Geomechanics and Engineering
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    • 제22권4호
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    • pp.329-337
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    • 2020
  • Polyacrylamide (PAM) possesses high water absorption capacity and a unique pH-dependent behavior that confer large potential to enhance the engineering performance of clays. In this study, calcium bentonite was treated with a nonionic PAM. Flexible-wall permeability test and the consolidation test were performed at different pH values to evaluate the effects of PAM treatment on the hydraulic and consolidation properties. Test results demonstrate that index properties are affected by the adsorbed PAM on clay surface: a decrease in specific gravity, a decrease in net zeta potential, and an increase in liquid limit are observed due to the PAM treatment. At a given pH, the compressibility of the treated clay is greater than that of the untreated clay. However, the compression indices of untreated and treated clays can be expressed as a single function of the initial void ratio, regardless of pH. Hydraulic conductivity is reduced by PAM treatment about 5 times at both neutral and alkaline pH conditions under similar void ratios, because of the reduction in size of the water flow channel by PAM expansion. However, at acidic pH, the hydraulic conductivity of the treated clay is slightly higher than the untreated clay. This reflects that the treated bentonite with PAM can be beneficially used in barrier system for highly alkaline residues.

Effect of Environmental pH on End Products, Fermentation Balances and Bioenergetic As-pects of Lactobacillus bulg-aricus in a Glucose-limited “pH Stat” Continuous Culture.

  • Rhee, Sang-Ki;Pack, Moo-Young
    • 한국미생물생명공학회:학술대회논문집
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    • 한국미생물생명공학회 1979년도 추계학술대회 심포지움
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    • pp.246.1-246
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    • 1979
  • A glucose-limited “pH-stat” continuous culture study of Lactobacillus bulgaricus NLS-4 in an anaerobic condition showed the marked effects of environmental pH on end products, fermentation blances and bioenergetic aspects of the organism. Lactic acid was the major end product of fermentation with minor products, such as acetic acid, formic acid and ethanol throughout the pH range tested. In acidic conditions below pH 6.5, a typi-cal pattern of homofermentation was revealed whereas in alkaline conditions, the metabolic pattern was changed from homofermentation to heterofermentation and led to acquire much energy. This metabolic change was likely due to the pH-dependent lactate dehydrogenase activity. Molar growth yields (Yglc=35.5-44.4) and YATP, $18.5\pm2.5$ in average which was 80% higher than the value ever postulated seemed to be accounted for less requirement of maintenance energy of the organism in the culture conditions.

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고분자전해질의 LbL multilayering 시 pH와 염 농도 조건에 따른 섬유의 전기화학적 특성 (Electrochemical Properties of Pulp Fiber with LbL Multilayering by Polyelectrolyte at the Different pH and Salt Concentration)

  • 류재호;이성린;진성민;윤혜정
    • 펄프종이기술
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    • 제40권4호
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    • pp.59-65
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    • 2008
  • Electrochemical properties of pulp fiber could be modified by LbL(Layer-by-Layer) multilayering. Salt concentration and pH affect the conformation and ionization degree of polyelectrolyte. In this study, therefore, we intended to evaluate the effect of pH and salt concentration in polyelectrolyte multilayering on the electrochemical properties of pulp fiber. The pH of the stock was varied to 4, 7, and 10. Salt concentration was controlled at 0.001, 0.003, and 0.005 M of NaCl. Higher zeta potential was obtained when pulp fibers were multilayered at the condition of higher pH and higher salt concentration. The final zeta potential of pulp fiber after multilayering was dependent on the adsorption ratio of polyelectrolyte and the initial zeta potential of fibers.

Drug Release from the Enzyme-Degradable and pH-Sensitive Hydrogel Composed of Glycidyl Methacrylate Dextran and Poly{acrylic acid)

  • Kim In-Sook;Oh In-Joon
    • Archives of Pharmacal Research
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    • 제28권8호
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    • pp.983-987
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    • 2005
  • Hydrogels composed of glycidyl methacrylate dextran (GMD) and poly(acrylic acid, PM) were prepared by UV irradiation method for colon-specific drug delivery. GMD was synthesized by coupling of glycidyl methacrylate to dextran in the presence of 4-(N,N-dimethylamino)pyridine. GMD was photo-polymerized by ammonium peroxydisulfate as initiating system in phosphate­buffered solution (0.1 M, pH 7.4). And then, acrylic acid monomer was added and subsequently heat-polymerized by 2,2'-azobisisobutyronitrile as an initiator. The hydrogels exhibited high swelling ratio (about 20) at $37^{\circ}C$, and showed a pH-dependent swelling behavior. In addition, the swelling ratio of the hydrogel was remarkably enhanced to about 45 times in the presence of dextranase at pH 7.4. The swelling-deswelling behavior proceeded reversibly for the GMD/PM hydrogels between pH 2 and pH 7.4. Release of 5-aminosalicylic acid from the GMD/PAA hydrogels was evaluated in simulated gastrointestinal pH fluids in the absence or presence of dextranase. We concluded that the hydrogels prepared could be used as a dual-sensitive drug carrier for sequential release in gastrointestinal tract.

Membrane-Bound Protease FtsH Protects PhoP from the Proteolysis by Cytoplasmic ClpAP Protease in Salmonella Typhimurium

  • Hyungkeun Song;Eunna Choi ;Eun-Jin Lee
    • Journal of Microbiology and Biotechnology
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    • 제33권9호
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    • pp.1130-1140
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    • 2023
  • Among the AAA+ proteases in bacteria, FtsH is a membrane-bound ATP-dependent metalloprotease, which is known to degrade many membrane proteins as well as some cytoplasmic proteins. In the intracellular pathogen Salmonella enterica serovar Typhimurium, FtsH is responsible for the proteolysis of several proteins including MgtC virulence factor and MgtA/MgtB Mg2+ transporters, the transcription of which is controlled by the PhoP/PhoQ two-component regulatory system. Given that PhoP response regulator itself is a cytoplasmic protein and also degraded by the cytoplasmic ClpAP protease, it seems unlikely that FtsH affects PhoP protein levels. Here we report an unexpected role of the FtsH protease protecting PhoP proteolysis from cytoplasmic ClpAP protease. In FtsH-depleted condition, PhoP protein levels decrease by ClpAP proteolysis, lowering protein levels of PhoP-controlled genes. This suggests that FtsH is required for normal activation of PhoP transcription factor. FtsH does not degrade PhoP protein but directly binds to PhoP, thus sequestering PhoP from ClpAP-mediated proteolysis. FtsH's protective effect on PhoP can be overcome by providing excess ClpP. Because PhoP is required for Salmonella's survival inside macrophages and mouse virulence, these data implicate that FtsH's sequestration of PhoP from ClpAP-mediated proteolysis is a mechanism ensuring the amount of PhoP protein during Salmonella infection.

자동 보정 주파수 의존형 근관장 측정기의 개발 (Development of a Frequency Dependent Type Apex Locator with Automatic Compensation)

  • 김덕원;남기창;김영주;이승종
    • 대한의용생체공학회:의공학회지
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    • 제19권6호
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    • pp.595-602
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    • 1998
  • 근관장 측정기 중 주파수 의존형이 다른 방법보다 정확도도 높고 사용이 간편한 것으로 평가되고 있다. 그러나 주파수 의존형 또한 실제 임상 사용시 근관내의 전해질 유무에 영향을 받는다. 본 연구에서는 전해질의 영향을 최소화한 자동 보정 주파수 의존형 전자 근관장 측정기를 개발하였다. 또한 500Hz와 10kHz가 임피던스 측정을 위해 사용되는 교류신호로써 측정오차를 최소화하는 최적 주파수임을 상품화된 제품의 주파수(400Hz, 8kHz)와 비교하여 확인하였으며, 신호의 파형에 따른 통계적 유의성은 없었다(p>0.05). 두 신호의 임피던스 비는 파일의 근관 내의 위치를 알려주고, 본 연구에서 적용한 두 신호의 전압차는 근관내 용액의 상태를 나타내 준다. 전압차를 이용하여 보정한 결과 측정오차를$H_2O_2$ 용액에서는 평균 +0.54mm에서 +0.18mm로(p<0.01), NaOCl 용액에서는 평균 -0.33mm에서 -0.01mm로(p<0.01) 줄일 수 있었다. 정확도는 허용오차를 ${\pm}$0.5mm를 기준으로 할 때, $H_2O_2$ 용액에서는 71.1%에서 91.1%로, NaOCl 용액에서는 82.2%에서 100%로 향상시켰다.

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