• Title/Summary/Keyword: Acidic proteins

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Fractionation of enzymatically methylated acid-insoluble proteins from thymus nuclei

  • Lee, Hyang-Woo;Kim, Sang-Duk;Paik, Woon-Ki
    • Archives of Pharmacal Research
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    • v.9 no.3
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    • pp.157-161
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    • 1986
  • Isolated calf thymus nuclei were in vitro methylated with S- adenosy-L-methyl-$^{14}C$ methionine, and the proteins were fractionated according to their solubilities. Histone fraction ($H_{2}SO_{4}$-soluble fraction) contained approximately 60% total radioactivity incorporated, while "residual protein" which was ($H_{2}SO_{4}$-insoluble contained the remaining radio-activity. The "residual protein" was further fractionated into various acidic proteins, which contained very littel of the radioactivity. However, the protein fraction eluted from DEAE-cellulose with 0.5 N NaOH contained the largest amount of radioactivity. This protein was found to be basic in nature by amino analysis.

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Protein Expression Analysis of Halobacillus dabanensis $D-8^T$ Subjected to Salt Shock

  • Feng De Qin;Zhang Bo;Lu Wei Dong;Yang Su Sheng
    • Journal of Microbiology
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    • v.44 no.4
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    • pp.369-374
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    • 2006
  • To investigate the mechanism of salt tolerance of gram-positive moderately halophilic bacteria, two-dimensional gel electrophoresis (2-D PAGE) was employed to achieve high resolution maps of proteins of Halobacillus dabanensis $D-8^T$. Approximately 700 spots of proteins were identified from these 2-D PAGE maps. The majority of these proteins had molecular weights between 17.5 and 66 kDa, and most of them were distributed between the isoelectric points (pI) 4.0 and 5.9. Some protein spots were distributed in the more acidic region of the 2-D gel (pI <4.0). This pattern indicated that a number of proteins in the strain $D-8^T$ are acidic. To understand the adaptation mechanisms of moderately halophilic bacteria in response to sudden environmental changes, differential protein profiles of this strain were investigated by 2-D PAGE and $Imagemaster^{TM}$ 2D Platinum software after the cells were subjected to salt shock of 1 to 25% salinity for 5 and 50 min. Analysis showed 59 proteins with an altered level of expression as the result of the exposure to salt shock. Eighteen proteins had increased expression, S proteins were induced, and the expression of 33 proteins was down-regulated. Eight of the up-regulated proteins were identified using MALDI-TOF/MS and MASCOT, and were similar to proteins involved in signal transduction, proteins participating in energy metabolism pathways and proteins involved in stress.

Analysis of Aluminum Stress-induced Differentially Expressed Proteins in Alfalfa Roots Using Proteomic Approach

  • Kim, Dong-Hyun;Lee, Joon-Woo;Min, Chang-Woo;Rahman, Md. Atikur;Kim, Yong-Goo;Lee, Byung-Hyun
    • Journal of The Korean Society of Grassland and Forage Science
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    • v.42 no.3
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    • pp.137-145
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    • 2022
  • Aluminum (Al) is one of the major factors adversely affects crop growth and productivity in acidic soils. In this study, the effect of Al on plants in soil was investigated by comparing the protein expression profiles of alfalfa roots exposed to Al stress treatment. Two-week-old alfalfa seedlings were exposed to Al stress treatment at pH 4.0. Total protein was extracted from alfalfa root tissue and analyzed by two-dimensional gel electrophoresis combined with MALDI-TOF/TOF mass spectrometry. A total of 45 proteins differentially expressed in Al stress-treated alfalfa root tissues were identified, of which 28 were up-regulated and 17 were down-regulated. Of the differentially expressed proteins, 7 representative proteins were further confirmed for transcript accumulation by RT-PCR analysis. The identified proteins were involved in several functional categories including disease/defense (24%), energy (22%), protein destination (9%), metabolism (7%), transcription (5%), secondary metabolism (4%), and ambiguous classification (29%). The identification of key candidate genes induced by Al in alfalfa roots will be useful to elucidate the molecular mechanisms of Al stress tolerance in alfalfa plants.

Protein Carboxylmethylation in Porcine Spleen is Mainly Mediated by Class I Protein Carboxyl O-Methyltransferase

  • Cho, Jae-Youl;Kim, Sung-Soo;Kwon, Myung-Hee;Kim, Seong-Hwan;Lee, Hyang-Woo;Hong, Sung-Youl
    • Archives of Pharmacal Research
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    • v.27 no.2
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    • pp.206-216
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    • 2004
  • The functional role of protein carboxylmethylation (PCM) has not yet been clearly elucidated in the tissue level. The biochemical feature of PCM in porcine spleen was therefore studied by investigating the methyl accepting capacity (MAC) of natural endogenous substrate proteins for protein carboxyl O-methyltransferase (PCMT) in various conditions. Strong acidic and alkaline-conditioned (at pH 11.0) analyses of the MAC indicated that approximately 65% of total protein methylation seemed to be mediated by spleen PCMT. The hydrolytic kinetics of the PCM products, such as carboxylmethylesters (CMEs), under mild alkaline conditions revealed that there may be three different kinds of CMEs [displaying half-times (T$_{1}$2/) of 1.1 min (82.7% of total CMEs), 13.9 min (4.6%), and 478.0 min (12.7%)], assuming that the majority of CME is base-labile and may be catalyzed by class I PCMT. In agreement with these results, several natural endogenous substrate proteins (14, 31 and 86 kDa) were identified strikingly by acidic-conditioned electrophoresis, and their MAC was lost upon alkaline conditions. On the other hand, other proteins (23 and 62 kDa) weakly appeared under alkaline conditions, indicating that PCM mediated by class II or III PCMT may be a minor reaction. The MAC of an isolated endogenous substrate protein (23-kDa) was also detected upon acidic-conditioned electrophoresis. Therefore, our date suggest that most spleen PCM may be catalyzed by class I PCMT, which participates in repairing aged proteins.

A study of ribonuclease activity in venom of vietnam cobra

  • Nguyen, Thiet Van;Osipov, A.V.
    • Journal of Animal Science and Technology
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    • v.59 no.9
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    • pp.20.1-20.9
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    • 2017
  • Background: Ribonuclease (RNase) is one of the few toxic proteins that are present constantly in snake venoms of all types. However, to date this RNase is still poorly studied in comparison not only with other toxic proteins of snake venom, but also with the enzymes of RNase group. The objective of this paper was to investigate some properties of RNase from venom of Vietnam cobra Naja atra. Methods: Kinetic methods and gel filtration chromatography were used to investigate RNase from venom of Vietnam cobra. Results: RNase from venom of Vietnam cobra Naja atra has some characteristic properties. This RNase is a thermostable enzyme and has high conformational stability. This is the only acidic enzyme of the RNase A superfamily exhibiting a high catalytic activity in the pH range of 1-4, with $pH_{opt}=2.58{\pm}0.35$. Its activity is considerably reduced with increasing ionic strength of reaction mixture. Venom proteins are separated by gel filtration into four peaks with ribonucleolytic activity, which is abnormally distributed among the isoforms: only a small part of the RNase activity is present in fractions of proteins with molecular weights of 12-15 kDa and more than 30 kDa, but most of the enzyme activity is detected in fractions of polypeptides, having molecular weights of less than 9 kDa, that is unexpected. Conclusions: RNase from the venom of Vietnam cobra is a unique member of RNase A superfamily according to its acidic optimum pH ($pH_{opt}=2.58{\pm}0.35$) and extremely low molecular weights of its major isoforms (approximately 8.95 kDa for RNase III and 5.93 kDa for RNase IV).

Electrophoretic pattern and classification of chitinolytic enzymes from Rehmannia glutinosa (지황 chitin 분해효소의 전기영동 양상 및 분류)

  • Pan, Cheol-Ho;Lee, Eun-A;Jo, Do-Hyun;Kim, Su-Il
    • Applied Biological Chemistry
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    • v.41 no.7
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    • pp.505-509
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    • 1998
  • Chitinase, which is one of the pathogenesis-related proteins, was examined from Rehmannia glutinosa. Rehmannia chitinases were classified into basic and acidic groups by chitin affinity column chromatography. According to the chitinase pattern by native PAGE, basic chitinases of low Rf values were extracted more under an acidic condition (pH 2.9) than a basic condition (pH 8.8), while acidic chitinases having high Rf values were mainly detected in basic extracts. There were in total seven chitinase isoforms of three basic and four acidic isoforms in Rehmannia. The range of molecular weight of Rehmannia chitinase was from 28 to 32 kD. It showed the possibility of tissue specific expression of acidic chitinases and basic chitinases were mainly detected in root.

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A Study on the Polymorphisms in Parotid Salivary Proteins of the Patients with Diabetes Millitus (당뇨병 환자의 이하선 타액내 단백질의 다형현상에 대한 연구)

  • Jong-Mo Ahn;Chang-Lyuk Yoon
    • Journal of Oral Medicine and Pain
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    • v.17 no.2
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    • pp.99-108
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    • 1992
  • The purpose of this study was to evaluate the polymorphosms in parotid salivary proteins of the patients with diabetes mellitus. Saliva from the parotid glands was collected from 94 healthy Korean adults who were live in Kwang-ju and from 33 diabetes mellitus patients who had more than 140mg/dl of fastingblood sugar for one week. Diabetes mellitus patient group was subdivided to insulin dependent diatetes mellitus (IDDM) and non-insulin dependent diabetes mellitus (NIDDM). In the saliva collected from the parotid glands, parotid acidic protein(Pa), proline-rich protein(Pr) and double band protein(Db) were analyzed to evaluate the distribution of phenotype using alkaline slab polyacrylamide gel electrophoresis. The results were as follows : 1. The parotid acidic protein (Pa) was found more frequently in the diabetes mellitus patient group than in the control group, but the difference was not statistically significant. 2. The Pr(1-2) type was found more frequently in the control group, but the Pr(1-1) and Pr(2-2) type were found more freqnently in the diabetes mellitus patient group and the difference of phenotypic distribution was statistically significant between the two groups. (p<0.05) 3. The parotid acidic protein(Pa) and Pr(1-2) type were found more frequently in the noninsulin dependent diabetes mellitus (NIDDM) patients than in the insulin dependent diabetes mellitus patients, though the difference was not statistically significant.

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Interaction between Whey and Soybean Proteins (유청 및 대두 단백질의 상호작용)

  • Shon, Dong-Hwa;Lee, Hyong-Joo
    • Applied Biological Chemistry
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    • v.31 no.4
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    • pp.361-370
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    • 1988
  • To investigate the interaction between whey and soybean protein, thermal changes of component proteins were analyzed by column chromatography and gel electrophoresis. In the Sephadex G-200 chromatography of the mixture treated at above $80^{\circ}C$, the amount of low molecular weight proteins and high molecular aggregates were increased. This implicated that dissociation of 1ls globulin into subunits and the formation of soluble aggregates between these subunits and whey proteins that contain thiol and disulfide groups. These interaction between soy proteins and ${\beta}-lactoglobulin$, ${\alpha}-lactalbumin$, and proteose-peptone 3 were confirmed by gel electrophoresis. Bovine serum albumin, Immunoglobulin-G(H), Lactoferrin, 1ls-subunits(basic and acidic), and subunit of 7s globulin were also considered to interact each other depending on the condition of the salt solutions.

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The Gene Frequency in Parotid Salivary PR, DB, PA Proteins according to Salivary Secretory Blood Components (타액내 혈형물질 분비형 및 비분비형에 따른 이하선 타액내 Pr,Db,Pa 단백질의 유전자 빈도)

  • San Kim;Chang-Lyuk Yoon
    • Journal of Oral Medicine and Pain
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    • v.19 no.2
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    • pp.233-244
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    • 1994
  • The purpose of this study was to evaluate the gene frequency in parotid salivary proteins according to salivary blood components and salivary blood types. Parotid and whole saliva were collected from 160 healthy Korean adults (from 20 years of age to 43). They were divided by blood type(Q,B, AB,O type). Each group contained 40 adults respectively. They were tested to the salivary secretory blood components and parotid acidic protein(Pa), proline-rich protein(Pr) and double band protein(Db) were analyzed to evaluate the distribution of phenotype using alkaline slab polyacrylamide gel electrophoresis. Results were as follows : 1. In parotid saliva, the salivary blood substances were not found. In whole saliva, secretory type was 21.9% and non-secretory type was 78.1%. : In A type blood group, secretory type 87.5% and non-secretory type 12.5%. In B type blood group, secretory type 82.5% and non-secretory type 17.5%. In AB type blood group, secretory type 85% and non-secretory type 15%. In O type blood group, secretory type 57.5% and non-secretory type 42.5%. 2. The gene frequency of parotid acidic protein(Pa) were Pa+=0.160, Pa-=0.840 and proline-rich protein(Pr) were Pr1=0.781, Pr2=0.219 and double-band protein(Db) were Db+=0.019, Db-=0.981. 3. The difference between phenotype of Pa, Pr, Db proteins and salivary secretory blood components was not statistically significant. (P>0.05) 4. The difference between phenotype of Pa, Pr, Db proteins and blood types was not statistically significant.(P>0.05)

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Bacterial Multiplications and Electrophoretic Patterns of Soluble Proteins in Compatible and Incompatible Interactions of Pepper Leaves with Xanthomonas campestirs pv. vesicatoria (Xanthomonas campestris pv. vesicatoria에 감염된 고추잎의 친화적, 불친화적 반응에서 세균증식과 수용성 단백질의 전기영동 패턴)

  • 이연경;김영진;황병국
    • Korean Journal Plant Pathology
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    • v.10 no.4
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    • pp.305-313
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    • 1994
  • Typically susceptible lesions were developed on pepper (cv. Hanbyul) leaves inoculated with the compatible strains Ds 1 of Xanthomonas campestris pv. vesicatoria. The lesions appeared first water-soaked and then turned yellow with a chlorotic area. In contrast, the leaves inoculated with the incompatible strain 81-23 initially turned yellow and then developed local necrosis. Multiplication of x. c. pv. vesicatoria in pepper leaves also were distinctly different between the two strains. The strain Ds 1 multiplied more greatly than did the strain 81-23 in the infected leaves. X. c. pv. vesicatoria infection of pepper leaves induced the synthesis of soluble proteins, especially more greatly in the compatible than in the incompatible interactions. Some pathogenesis-related (PR) proteins were detected in the intercellular washing fluid (IWF) and extracts of the infected pepper leaves. In particular, the 32 kDa protein on SDS-PAGE gels appeared intensely in the incompatible interaction. In contrast, some proteins with moluecular masses of 65, 71, and 75 kDa disappeared in the infected pepper leaves. Isoelectric focusing could identify the pIs of soluble proteins in infected pepper leaves. The accumulation of the IWF from infected leaves was more conspicuous in the incompatible than the compatible interaction. These results suggest that some extremely acidic and basic proteins were induced and accumulated in the intercellular spaces of infected pepper leaves.

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