• Title/Summary/Keyword: Sulfhydryl groups

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The changes in the Amounts of SH Compounds in Chlorella during the Synchronous Culture (Chlorella의 생활사를 통한 -SH화합물 함량의 변화)

  • 최호형;이영녹
    • Korean Journal of Microbiology
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    • v.19 no.1
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    • pp.8-13
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    • 1981
  • The content of sulfhydryl compounds in Chlorella cells during the life cycle in the synchronous culture is determined spectrophotomatically at 250nm(pH7.0) using p-CMB as SH-reagent. The changes in the content of-SHl compounds and protein in Chlorella cells is measured during the life cycle in connection with cell division and analyzed. 1) The amounts of total ninhydrin reactive substance increased with growth of cells but increased the more at the $L_4$ stage(cytokinesis stage) than at the $L_2$ stage (nuclear division stage). 2) The sulfhydryl content of Chlorella cells increased strikingly at the $L_2$ stage and decreased markedly at the $L_4$ stage. 3) The amounts of values -SH/protein showed a peak at the $L_2$ stage. The increase of the amount of total-SH compounds of cells during the nuclear division period was considered to be caused by the weighty roles of protein-SH groups for the formation of nuclear division apparatus and for the enzyme activity.

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Changes in Functional Groups of Protein by Lipid Deterioration in the Biological System of Rice Bran (미강 저장 중 지방의 산패에 따라 생성된 산화 지질이 단백질의 기능기에 미치는 영향에 관한 연구)

  • Song, Yeong-Ok;Choi, Hong-Sik
    • Korean Journal of Food Science and Technology
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    • v.22 no.5
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    • pp.596-601
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    • 1990
  • The effects of peroxidized lipid on the protein in the biological system of rice bran was studied by determining the changes in the content of functional groups under two different storage conditions. One stored at controlled atmosphere of $35^{\circ}C$ with relative humidity 65% and the other one was exposed to the air of $25^{\circ}-30^{\circ}C$ with relative humidity 70-90%. The lipid peroxidation started after the lipolysis was almost completed. The autoxidation occurred much faster in the bran exposed to the air than that stored in the controlled atmosphere. Substantial changes in the physiochemical characteristics were observed in all of the major functional groups in both of the samples. The content of sulfhydryl and available lysine decrease·1 as lipid peroxidation progressed. Protease activity was lost almost completely. Protein solubility and in vitro digestibility also decreased during storage. The lipid peroxidation and contents of major protein functional groups were significantly correlated (p<0.05) and the correlation coefficients were higher than -0.8, for the both of the sample. peroxidized lipid was found to deteriorate protein in the biological system as well.

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Myosin Heavy Chain Covalenily Modified at Its Reactive Site Sulfhydryl Residues is Preferentially Degraded by Calpain (활성화 부위의 황화기가 화학적으로 변형된 Myosin Heavy Chain의 Calpain에 의한 선택적 분해)

  • 곽규봉;정성수;이창호;하두봉;정진하
    • The Korean Journal of Zoology
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    • v.36 no.3
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    • pp.347-352
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    • 1993
  • N-ethylmaleimide at low concentrations is known to interact specifically with 2 sulfhydryl residues in myosin heavy chain. Calpain, a Ca$^2$+-dependent neutral protease isolated from chick skeletal muscle, was found to preferentially degrade the alkylated protein but much less significantly the unmodified protein. Exposure of myosin to KMnO$_4$, which is also known to interact with sulthydryl groups, also caused the rapid degradation of the myosin heavy chain. Furthermore, treatment of each agent with increasing concentrations results in a greater loss of the myosin ATPase activity, indicating that the modification occurred at the reactive site sulfhydryl residues. These results suggest that the covalent modification at the reactive site salfhydryl residues in the myosin heavy chain may mark the protein for degradation by intracellular proteases such as calpain.

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Structural Arrangement for Functional Requirements of Brain Recombinant 4-Aminobutyrate Aminotransferase

  • Sung, Bo-Kyung;Kim, Young-Tae
    • BMB Reports
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    • v.33 no.1
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    • pp.43-48
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    • 2000
  • 4-Aminobutyrate aminotransferase is a key enzyme of the 4-aminobutyric acid shunt. It converts the neurotransmitter 4-aminobutyric acid to succinic semialdehyde. In order to study the structural and functional aspects of catalytically active Cys residues of pig brain 4-aminobutyrate aminotransferase, we purified the active form in E. coli by coproduction of thioredoxin. The structural arrangement for functional requirements of a dimeric protein using a bifunctional sultbydryl reagent was then characterized, and the spatial proximity between the essential SH groups and a cofactor (pyridoxal-5'-phosphate) binding site was determined. The bifunctional sultbydryl reagent DMDS reacted with the enzyme at the ratio of one molecule per enzyme dimer. This resulted in an approximately 50% loss of enzymatic activity. The spatial proximity of the distance between the essential SH groups and the cofactor-binding site was determined by the energy transfer measurement technique. The result (approximate 20 ${\AA}$) suggested that cross-linking of two sulfhydryl groups with DMDS is not near a PLP binding site.

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Effect of Single or Fractionated X-Irradiation on the NP-SH and NP-SS of Rabbit Liver and Lung Tissues (단회(單回) 및 분획(分劃) X선(線) 조사(照射)가 가토(家兎)의 간(肝) 및 폐조직(肺組織)의 NP-SH 및 NP-SS에 미치는 영향(影響))

  • Park, Soo-Ha;Choi, Kyung-Sook;Kim, Byung-Kwon;Kwon, Yong-Ju;Lee, Jae-Bok
    • The Korean Journal of Physiology
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    • v.4 no.2
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    • pp.61-67
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    • 1970
  • In view of the importance of sulfhydryl groups in producing a chemical protection against ionizing radiation, an attempt was made to evaluate the changes of intrinsic non-protein sulfhydryl (NP-SH) and non-Protein disulfide(NP-SS) of the lung and liver tissues of rabbits following the whole body X-irradiation with 900 r either in single or fractionated $(300\;r{\times}3)$ dose. NP-SH was measured by Ellman's method, and NP-SS was measured by the electrolytic reduction method described by Dohan. Experiment was performed at 1,3,5,24 and 48 hours post·irradiation, and the results were compared with the control. The results thus obtained are summarized as follows; 1) Intrinsic levels of NP-SH and NP-SS of normal rabbits were $0.77{\pm}0.10$ and $0.61{\pm}0.07\;{\mu}mol/gm$ wet weight in liver, and $0.28{\pm}0.03$ and $0.54{\pm}0.03$ in lung tissues respectively. 2) NP-SH of liver after single X-irradiation showed no significant change in general, but at 48 hours post·irradiation, it was elevated comparing with the normal value. 3) Levels of NP-SS in liver was decreased than the normal value in the irradiated groups, and the lowest level was observed at 3 hours after single X-irradiation and at 5 hours after fractionated irradiation. 4) In lung tissues, levels of NP-SS showed no significant change from the control at earlier experimental hours, but a great decrease was observed at later Part of the experiment.

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The Effects of Onion and Garlic on Copper-Phenanthroline Complex Induced DNA Degradation (Copper-Phenanthroline 복합체에 의해 유도되는 DNA 손상에 대한 양파와 마늘의 억제효과)

  • 박평심;이명렬
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.21 no.4
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    • pp.367-371
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    • 1992
  • In the present study, the effects of onion and garlic on copper and 1, 10-phenanthroline com plex induced DNA degradation were investigated by the decreased level of thiobarbiturin acid (TBA) reactive materials. Phenanthroline is specific for copper and the reaction releases TBA reactive material from DNA which can be measured by absorbance at 535nm. The levels of TBA reactive materials were decreased by adding onion or garlic ghomogenate into reaction mixture but the onion had more strong potency and the effect of onion was not changed by boiliing. Superoxide dismutase (SOD) and catalase have no inhibitory effects on copper induced DNa damage but reduced glutathone was more effective. The activities of antioxidant enzymes and the contents of sulfhydryl groups in onion and garlic were also investigated. The activity of SOD was more higher in garlic, but catalase and glutathione peroxidase activities were higher in onion. The contents of induced DNA damage were not by antioxidant enzymes such as SOD, catalase and glutathione peroxidase or sulfhydryl groups, but a substance which is more stable in high temperature.

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The Physicochemical Properties of Modified Soybean Protein Isolate by Dimethylglutarylation (Dimethylglutarylation에 의한 변형대두단백질의 물리화학적 특성)

  • Choi, One-Kyun;Jung, Chul-Won
    • Journal of the Korean Society of Food Culture
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    • v.14 no.5
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    • pp.467-476
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    • 1999
  • We studied to improve the functional properties of soybean protein isolate by dimethylglutarylation and acetylation. Soybean protein isolate was acylated rapidly up to 80% modification and more of 80% modification was proceed slowly. Electrophoretic analysis showed that more changes in modified protein. Also, modification of soybean protein produced more ionizable tyrosines and exposed more hydrophobic groups, while modified protein exhibited a loss of reactive sulfhydryl groups. Spectrophoretic studies demonstrated that the shift was occurred at the UV 278nm and fluorescence 333nm, respectively, and the intensity decreased as the degree of modification was increased.

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Characterization of Cysteine Residues in Cabbage Phospholipase D by Sulfhydryl Group Modifying Chemicals (설프히드릴 변형 화합물질들에 의한 양배추 포스포리파제 D의 시스테인 잔기의 특성)

  • Go, Eun-Hui
    • Journal of the Korean Chemical Society
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    • v.50 no.5
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    • pp.362-368
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    • 2006
  • SH group modifying chemicals were used to characterize the eight cysteine residues of cabbage PLD. 5,5-dithiobis(2-nitrobenzoate)(DTNB) was used to titrate the SH group of cysteine residues . Based on the optical density at 412nm due to the reduced DTNB, 4 SH groups are found to be present in a native PLD while 8 SH groups in the denatured PLD whose tertiary structure was perturbed by 8M urea. The results imply that among the 8 cysteine residues of PLD, the half(4) are exposed on the surface whereas the other half are present at the interior of the enzyme tertiary structure. The PLD was inactivated by SH modifying reagents such as p-chloromercuribenzoate(PCMB), iodoacetate, iodoacetamide, and N-ethylmaleimide. At the addition of dithiothreitol(DTT) only the PCMB inhibited PLD activity was recovered reversibly. The micro-environment of the exposed SH group of cysteine residues was examined with various disulfide compounds with different functional groups and we found that anionic or neutral disulfides appear to be more effective than the positively charged cystamine for inactivating the PLD activity. The effect of redox state of cysteine residues on the PLD activity was further explored with H2O2. The oxidation of SH groups by H2O2 inhibited the PLD activity more than 70%, which was mostly recovered by DTT. From these results, we could confirm chemically that all the cysteine residues of PLD are present as in their reduced SH forms and the 4 SH groups exposed on the surface of the enzyme may play important roles in the regulation of PLD activity.

Effect of X-Irradiation on the Levels of some Sulfhydryl Groups, Protein and Cell Volume of Ehrlich Ascites Tumour Cells (X-선(線) 조사(照射)가 Ehrlich 암세포(癌細胞)의 용적(容積), 단백양(蛋白量) 및 수종(數種) Sulfhydryl 기(基)에 미치는 영향(影響)에 관(關)하여)

  • Yu, Choon-Shik;Choo, Young-Eun
    • The Korean Journal of Physiology
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    • v.3 no.2
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    • pp.9-16
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    • 1969
  • It is well known that a number of -SH and -SS containing substances afford a certain measure of protection against radiation effects in many biological systems, and it is conceivable that inherent -SH levels in Ehrlich ascites tumour (ELD)cells may be of decisive improtance with respect to the development of cellular radiation injury. So far, little effort has been directed to elucidate the changes in levels of different -SH and -SS groups in ELD cells when the tumour-bearing whole animal was subjected to the sublethal dose of X-irradiation. The present study was designed to bring some lights in the possible changes of and relationship between various sulfhydryl levels, such as P-SH, NP-SH and NP-SS, as well as the content of protein and cell volume of ELD cells, after subjecting the ELD mice to 1,200 r of X-irradiation. The animals used in this experiment were all mixed bred mice of $20{\sim}25\;gm$ in body weight (approximately 2 months old) irrespective of sex. 12 mice in one experiment were inoculated intraperitoneally with 0.2 ml of ascites tumour cells $(2{\times}10^6\;cells)$, and on the 7th day of the tumour growth, they were X-irradiated with 1,200 r, using the conventional X-ray machine under the following conditions: 200 Kv at 15 mA, 0.5 mm Cu filter, target-skin distance: 50 cm. Radiation dose was measured with the the Philip integrating dosimeter. At 24, 36, 48 and 60 hours after the X-irradiation, the mice were killed by cervical dislocation, and the tumours were taken out. Freshly withdrawn ascites tumours were placed in ice, and immediately the cell concentration was measured with the Coulter Cell Counter (Model B), and the hematocrit of the tumour cells were also determined. Cell volume was thus calculated by the cell concentration and hematocrit value. P-SH content of ELD cells was measured potentiometrically according to the method of Calcutt & Doxey, and NP-SH and NP-SS contents were measured spectrophotometrically by the method described by Ellman. Protein content of ELD cells was determined with the Folin phenol reagent by Lowry et al. Altogether, 48 experimental mice were used, and 12 mice with the only exception of X-irradiation were used as the control. Results obtained indicate that the contents of all the cellular sulfhydryl groups as well as cell volume and protein content of the ELD cells increase significantly as time progresses after the sub-lethal X-ray dose of 1,200 r was given and that all the increase is in a lineal fashion. The regression lines of the relative values, (i. e., taking each control value as 1) of all the values obtained, and the regression lines of cell volume, protein and NP-SH are identical, whereas those of NP-SS and P-SH appear to be widely seperated. However, the difference of those two lines (NP-SS & P-SH) were found to be not significant statistically (p>0.05). Therefore, it can be concluded from the above results that all the values examined increase in a lineal fashion with no statistically significant difference among them. Also, with the radiation dose of 1,200 r, the ELD cell becomes enlarged and swollen progressively up to 60 hours post-irradiation and it becomes more than two times of the original normal size at 60 hours after the irradiation, and up to this stage, it seems apparent that the cell division has been slow due to the X-irradiation applied in this experiment. It is well understandable that the contents of NP-SH, NP-SS, P-SH and protein of the ELD cells increase in parallel with the increase of the cell volume by the X-ray does used, but it also seems interesting to note that all the cellular substances tested show no appreciable difference in the pattern of increase.

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Changes of Indicative Substances According to Heat Treatment of Milk (우유의 가열처리에 따른 지표물질의 변화)

  • 김경미;홍윤호;이용규
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.21 no.4
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    • pp.390-397
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    • 1992
  • This study was carried out to analyze the physicochemical properties of bovine milks, which were heated with LTLT, HTST, UHT pasteurization and UHT sterilization methods and to compare the heat intensity among the heating methods and samples. The mean HMF values per liter milk were measured as 0.66~1.62 $\mu$M (LTLT), 0.9~1.78$\mu$M (HTST), 3.53$\mu$M(UHT pasteurized) and 7.43~8.97$\mu$M (UHT sterilized) in samples, re- sportively. The available Iysine contents per 100ml milk showed 293.2 mg (Raw), 289.2~291.2 mg (LTLT), 298.4~292.4mg (HTST), 272.4~261.6mg (UHT pasteurized) and 279.0mg (UHT sterilized), respectively. The rates of whey protein denaturation were 9.5~11.4% (LTLT), 9.5~17.1% (HTST), 89.3~95% (UHT pas-tsterilized) and 62.7% (UHT sterilized), respectively. The contents of SH groups per g protein were determined as 2.86$\mu$M (Raw) and 2.95~3.15$\mu$M (LTLT), 3.08~3.18$\mu$M (HTST), 3.26~3.42$\mu$M (UHT Pasteurized) and 3. 36$\mu$M (UHT sterilized), respectively, The SS groups Contents per g protein were 28.93$\mu$M (Raw), 25.72~26. 51 $\mu$M (LTLT), 26.93~26.79$\mu$M (HTST), 23.65~23.04 $\mu$M (UHT pasteurized) and 24.69$\mu$M (UHT sterilized), respectively. The ascorbic acid contents per liter milk were measured 6.05mg (Raw), 1.47~1.65mg (LTLT), 2.50~3.85mg (HTST), 2.87~3.69mg (UHT pasteurized) and 4.50mg (UHT sterilized). The changes of some in-dices in milk samples depend on the heating temperature and time ; the HMF values, SH groups, whey protein denaturation rates increased, while the available lysine contents and SS groups decreased in LTLT, HTST, UHT pasteurized and UHT sterilized milks. No remarkable differences were found in heating indicators between LTLT and UHT milks.

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