• Title/Summary/Keyword: thermal denaturation.

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"OPEN" STRUCTURE OF SecA PROTEIN OF ESCHERICHIA COLI IN SOLUTION

  • Maengseok Song;Kim, Hyounman
    • Proceedings of the Korean Biophysical Society Conference
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    • 1996.07a
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    • pp.27-27
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    • 1996
  • SecA protein which has a pivotal role in the preprotein cranslocation across the inner membrane of Escherichia coli is a water-soluble protein with an unusual property of penetrating the membrane readily. An interesting and important question is what structural characteristics of SecA enables its ready penetration of lipid bilayer. The conformational properties of SecA in solution at 3$0^{\circ}C$, pH 7.5 were observed by hydrogen-tritium (HT) exchange, and denaturant-induced denaturation/renaturation and thermal unfolding. (omitted)

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Fabrication, Estimation and Trypsin Digestion Experiment of the Thermally Isolated Micro Teactor for Bio-chemical Reaction

  • Sim, Tae-Seok;Kim, Dae-Weon;Kim, Eun-Mi;Joo, Hwang-Soo;Lee, Kook-Nyung;Kim, Byung-Gee;Kim, Yong-Hyup;Kim, Yong-Kweon
    • JSTS:Journal of Semiconductor Technology and Science
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    • v.5 no.3
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    • pp.149-158
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    • 2005
  • This paper describes design, fabrication, and application of the silicon based temperature controllable micro reactor. In order to achieve fast temperature variation and low energy consumption, reaction chamber of the micro reactor was thermally isolated by etching the highly conductive silicon around the reaction chamber. Compared with the model not having thermally isolated structure, the thermally isolated micro reactor showed enhanced thermal performances such as fast temperature variation and low energy consumption. The performance enhancements of the micro reactor due to etched holes were verified by thermal experiment and numerical analysis. Regarding to 42 percents reduction of the thermal mass achieved by the etched holes, approximately 4 times faster thermal variation and 5 times smaller energy consumption were acquired. The total size of the fabricated micro reactor was $37{\times}30{\times}1mm^{3}$. Microchannel and reaction chamber were formed on the silicon substrate. The openings of channel and chamber were covered by the glass substrate. The Pt electrodes for heater and sensor are fabricated on the backside of silicon substrate below the reaction chamber. The dimension of channel cross section was $200{\times}100{\mu}m^{2}$. The volume of reaction chamber was $4{\mu}l$. The temperature of the micro reactor was controlled and measured simultaneously with NI DAQ PCI-MIO-16E-l board and LabVIEW program. Finally, the fabricated micro reactor and the temperature control system were applied to the thermal denaturation and the trypsin digestion of protein. BSA(bovine serum albumin) was chosen for the test sample. It was successfully shown that BSA was successfully denatured at $75^{\circ}C$ for 1 min and digested by trypsin at $37^{\circ}C$ for 10 min.

Thermal-Denaturation of File Fish Myofibrillar Protein and Protective Effect of Sucrose, Sorbitol and Amino Acids (말쥐치 근원섬유단백질의 열안정성과 및 가지 첨가제의 영향)

  • CHOI Young-Joon;PYEUN Jae-Hyeung
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.18 no.5
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    • pp.455-463
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    • 1985
  • Thermal-denaturation of myofibrillar protein of dorsal skeletal muscle from file fish was investigated by measuring denaturation constant($K_D$) and thermodynamic parameters at various temperatures. The protective effects of sucrose, sorbitol and amino acids when added individually or combined were also discussed. The denaturation rate as reflected in inactivation of myofibrillar protein Ca-ATPase was followed the first order reaction. The $K_D$ values at $25^{\circ}C,\;30^{\circ}C,\;and\;35^{\circ}C$ were $19.52{\times}10^{-5},\;112.25{\times}10^{-5},\;and\;247.20{\times}10^{-5}$, respectively. The activation energy of the reaction at $30^{\circ}C$ was 43 kcal/mole. The protective effects of sucrose, sorbitol, glycine, alanine and Na-glutamate were increased with the concentration but the effects of sorbitol and Na-glutamate decreased beyond 1.0 mole. Basic amino acids such as arginine and lysine did not revealed any protective effect on the thermal denaturation. In case of mixed addition, the effects of Na-glutamate to glycine, sorbitol to glycine, and sorbitol to sucrose or sorbitol to Na-glutamate were enhanced 1.2 to 7.0 times as much as that of control (ratio of mixing; 1:1, range of concentration; 0.5 to 1.25 mole). Under the frozen condition at $-20^{\circ}C$, two mixtures such as Na-glutamate to glycine and sorbitol to sucrose apparently revealed the protective effects.

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Analysis of Red Pepper (Capsicum annuum) Genome (고추의 게놈 분석)

  • 안정선
    • Journal of Plant Biology
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    • v.39 no.1
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    • pp.57-61
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    • 1996
  • The genome of red pepper was investigated by thermal denaturation, reassociation kinetics and measurement of nuclear volume for its base composition, spectrum of kinetic components and genome size. Base composition was estimated to be 37% (G+C) based on melting temperature. The reassociation of 300 nt fragments analyzed by hydroxyapatite chromatography revealed the presence of three kinetic components differing in fraction of genome, kinetic complexity and number of copies as follows; 4.8% (fast) with $5.6{\times}10^{4}\;bp$ and 10,754, 26% (intermediate) with $1.9{\times}10^{6}\;bp$ and 177, and 65% (slow) with $8.48{\times}10^{8}\;bp$ and 1. These measurements demonstrate that the genome of red pepper has a 1C DNA content of $1.25{\times}10^{9}\;bp$, which is about 33% of $4.05{\times}10^{9}\;bp$ calculated from nuclear volume of $62.4\;\mu\textrm{m}^3/1C$..

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Comparative Study on the Postmortem Proteolysis and Shear Force during Aging of Pork and Beef Semitendinosus Muscles

  • Seokhee Han;Kyung Jo;Seul-Ki-Chan Jeong;Hayeon Jeon;Soeun Kim;Minkyung Woo;Samooel Jung;Seonmin Lee
    • Food Science of Animal Resources
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    • v.44 no.5
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    • pp.1055-1068
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    • 2024
  • The differences in the proteolytic patterns and shear force of pork and beef during aging were evaluated. Pork and beef semitendinosus muscles were obtained at 24 and 48 h postmortem, respectively, and aged at 4℃ for 0 (Day 0), 7 (Day 7), and 14 days (Day 14). Changes in the electrical conductivity were observed in pork on Day 7 and beef on Day 14. The calpain activity increased in pork (p<0.05) after 14 days of aging, whereas that of beef decreased on Day 7 (p<0.05). The cathepsin B activity in pork and beef increased between Day 7 and 14 (p<0.05). The content of α-amino group in the 10% trichloroacetic acid-soluble fraction increased between Day 7 and 14 in pork (p<0.05), but increased steadily in beef throughout aging (p<0.05). The electrophoretogram of the myofibrillar proteins revealed a 30 kDa protein band only in the beef lane on Day 14. The cooked pork had no significant changes in the shear force during aging periods (p>0.05), while the gradual decrease in the shear force with the increasing aging periods was shown in the cooked beef (p<0.05). Circular dichroism analysis of myosin extracts from pork and beef revealed thermal denaturation temperatures of 55℃ and 58℃, respectively. This study highlights the different post-mortem proteolytic patterns and thermal denaturation temperatures of myosin in pork and beef semitendinosus muscles, which contribute to distinct changes in the shear force during aging between pork and beef.

Thermal Design of PCR Chip for LOC (랩온어칩을 위한 중합효소 연쇄반응 칩의 열설계)

  • Kim, Deok-Jong;Kim, Jae-Yun;Park, Sang-Jin;Heo, Pil-U;Yun, Ui-Su
    • 연구논문집
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    • s.33
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    • pp.17-25
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    • 2003
  • In this work, thermal design of a PCR chip for LOC is systematically conducted. From the numerical simulation of a PCR chip based on the finite volume method, how to control the average temperature of a PCR chip and the temperature difference between the denaturation zone and the annealing zone is presented. The average temperature is shown to be controlled by adjusting heat input and a cooler as well as a heater is shown to be necessary to obtain three individual temperature zones for polymerase chain reaction. To reduce the time required, a heat sink for the cooler is not included in the calculation domain for the PCR chip and heat sink design is conducted separately by using a compact modeling method, the porous medium approach.

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Screening Molecular Chaperones Similar to Small Heat Shock Proteins in Schizosaccharomyces pombe

  • Han, Jiyoung;Kim, Kanghwa;Lee, Songmi
    • Mycobiology
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    • v.43 no.3
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    • pp.272-279
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    • 2015
  • To screen molecular chaperones similar to small heat shock proteins (sHsps), but without ${\alpha}$-crystalline domain, heat-stable proteins from Schizosaccharomyces pombe were analyzed by 2-dimensional electrophoresis and matrix assisted laser desorption/ionization time-of-flight mass spectrometry. Sixteen proteins were identified, and four recombinant proteins, including cofilin, NTF2, pyridoxin biosynthesis protein (Snz1) and Wos2 that has an ${\alpha}$-crystalline domain, were purified. Among these proteins, only Snz1 showed the anti-aggregation activity against thermal denaturation of citrate synthase. However, pre-heating of NTF2 and Wos2 at $70^{\circ}C$ for 30 min, efficiently prevented thermal aggregation of citrate synthase. These results indicate that Snz1 and NTF2 possess molecular chaperone activity similar to sHsps, even though there is no ${\alpha}$-crystalline domain in their sequences.

Fabrication of a polymerase chain reaction micro-reactor using infrared heating

  • Im, Ki-Sik;Eun, Duk-Soo;Kong, Seong-Ho;Shin, Jang-Kyoo;Lee, Jong-Hyun
    • Journal of Sensor Science and Technology
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    • v.14 no.5
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    • pp.337-342
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    • 2005
  • A silicon-based micro-reactor to amplify small amount of deoxyribonucleic acid (DNA) has been fabricated using micro-electro-mechanical systems (MEMS) technology. Polymerase chain reaction (PCR) of DNA requires a precise and rapid temperature control. A Pt sensor is integrated directly in the chamber for real-time temperature measurement and an infrared lamp is used as external heating source for non-contact and rapid heating. In addition to the real-time temperature sensing, PCR needs a rapid thermocycling for effective PCR. For a fast thermal response, the thermal mass of the reactor chamber is minimized by removal of bulk silicon volume around the reactor using double-side KOH etching. The transparent optical property of silicon in the infrared wavelength range provides an efficient absorption of thermal energy into the reacting sample without being absorbed by silicon reactor chamber. It is confirmed that the fabricated micro-reactor could be heated up in less than 30 sec to the denaturation temperature by the external infrared lamp and cooled down in 30 sec to the annealing temperature by passive cooling.

Temperature Distributions of the Lumbar Intervertebral Disc during Laser Annuloplasty : A Cadaveric Study

  • Lee, Min Hyung;Kim, Il Sup;Hong, Jae Taek;Sung, Jae Hoon;Lee, Sang Won;Kim, Daniel H.
    • Journal of Korean Neurosurgical Society
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    • v.59 no.6
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    • pp.559-563
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    • 2016
  • Objective : Low back pain, caused intervertebral disc degeneration has been treated by thermal annuloplasty procedure, which is a non-surgical treatement. The theoretical backgrounds of the annuloplasty are thermal destruct of nociceptor and denaturization of collagen fiber to induce contraction, to shrink annulus and thus enhancing stability. This study is about temperature and its distribution during thermal annuloplasty using 1414 nm Nd : YAG laser. Methods : Thermal annuloplasty was performed on fresh human cadaveric lumbar spine with 20 intact intervertebral discs in a $37^{\circ}C$ circulating water bath using newly developed 1414 nm Nd : YAG laser. Five thermocouples were attached to different locations on the disc, and at the same time, temperature during annuloplasty was measured and analyzed. Results : Thermal probe's temperature was higher in locations closer to laser fiber tip and on lateral locations, rather than the in depth locations. In accordance with the laser fiber tip and the depth, temperatures above $45.0^{\circ}C$ was measured in 3.0 mm depth which trigger nociceptive ablation in 16 levels (80%), in accordance with the laser fiber end tip and laterality, every measurement had above $45.0^{\circ}C$, and also was measured temperature over $60.0^{\circ}C$, which can trigger collagen denaturation at 16 levels (80%). Conclusion : When thermal annuloplasty is needed in a selective lesion, annuloplasty using a 1414 nm Nd : YAG laser can be one of the treatment options.

Studies on the Thermal Stability and Color of Free Drip released from Pork Muscle with pH, Concentration of NaCl and Phosphate (pH, NaCl 및 phosphate 첨가에 따른 돈육 드립의 열안정성 및 색에 관한 연구)

  • Kim, Cheon-Jei;Lee, Chang-Hyun;Song, Min-Seok;Lee, Eui-Soo;Cho, Jin-Kook
    • Korean Journal of Food Science and Technology
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    • v.32 no.6
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    • pp.1285-1290
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    • 2000
  • The objective of this study was to evaluate the characteristics on the thermal denaturation of free drip released from pork loin during chilled storage using DSC (differential scanning calorimetry) with pH, concentration of NaCl and phosphate. The increasing of pH stabilized the heat resistance of the proteins in drip. A $T_1$ greatly increased of $T_{max}$ by $6.33^{\circ}C$ incline from pH 5.5 to 6.5. And increasing the concentration of NaCl destabilized the heat resistance of drip. $T_1$ showed the greatest reduction of $T_{max}\;(9.41^{\circ}C)$ in the presence of 5% NaCl. The presence of STPP (Sodium Tripolyphosphate) enchanced the thermal stability of pork drip by $5.84^{\circ}C$ in the presence of 0.5% STPP. As temperature increased from 40 to $100^{\circ}C$, lightness $(L^*)$ increased from 41.1 to 69.5, while redness $(a^*)$ decreased from 26.70 to 5.40. Particularly, both values of $L^*-$ and $a^*-$ greatly changed by 78% from 40 to $60^{\circ}C$.

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