• Journal of Plant Biotechnology
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• 제35권3호
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• pp.185-193
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• 2008

We evaluated the response to salt stress of two different ginseng lines, STG3134 and STG3159, which are sensitive and tolerant, respectively, to salt treatment. Plants were exposed to a 5 dS/m salt solution, and chlorophyll fluorescence was measured. STG3134 ginseng was more sensitive than STG3159 to salt stress. To characterize the cellular response to salt stress in the two different lines, changes in protein expression were investigated using a proteomic approach. Total protein was extracted from detached salt-treated leaves of STG3134 and STG3159 ginseng, and then separated by two-dimensional polyacrylamide gel electrophoresis(2-DE). Approximately 468 protein spots were detected by 2-DE and Coommassie brilliant blue staining. Twenty-two proteins were found to be reproducibly up- or down-regulated in response to salt stress. Among these proteins, twelve were identified using MALDI-TOF MS and ESI-Q-TOF and classified into several functional groups: photosynthesis-related proteins(oxygen-evolving enhancer proteins 1 and 2, rubisco and rubisco activase), detoxification proteins(polyphenol oxidase) and defense proteins($\beta$-1,3-glucanase, ribonuclease-like storage protein, and isoflavone reductase-like protein). The protein levels of ribonuclease-like storage protein, which was highly induced in STG3159 ginseng as compared to STG3134, correlated tightly with mRNA transcript levels, as assessed by reverse-transcription(RT)-PCR. Our results indicate that salinity induces changes in the expression levels of specific proteins in the leaves of ginseng plants. These changes may, in turn, playa role in plant adaptation to saline conditions.

• 한국환경농학회지
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• 제32권1호
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• pp.48-54
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• 2013

수수 종자의 품종 간 특이적으로 발현하는 단백질을 동정하여 기능성 유전자를 확보하고 이들 유전자를 이용하여 수수의 기능성 강화 및 품종 판별 기술 개발을 위한 유용 유전자를 확보하고자 프로테오믹스 기법을 이용하여 수수 종자로부터 단백질을 추출하였다. 추출한 단백질을 이차원전기영동후, colloidal CBB 염색을 통해 품종 별로 발현에 차이를 보이는 단백질을 분석하였다. 총 652 개의 spot들 중에 8개의 단백질 spot들이 발현 정도에 변화를 보였으며, 이들 단백질을 MALDI-TOF/TOF MS와 MASCOT database를 통해 동정한 결과, RNA metabolism (spot 1, spot 4) HSP (spot 2), 저장 단백질 (spot 3, spot 5, spot 6), 산화-환원 (spot 8) 관련 단백질 등이 동정되었다. 특히 동정된 단백질은 주로 흰찰수수 (WCS)에서 발현 정도가 높게 나오는 경향을 보였으며, 흰찰수수 (WCS)에서 유일하게 발현 되는 단백질로 Cupin family protein, Gloubulin 등이 동정되었다. DEAD-box helicase는 흰찰수수 (WCS)를 제외한 나머지 세 품종에서 발현되었다. Ribonuclease T2와 Aldo-Keto reductase는 대풍수수 (DPS)를 제외한 나머지 세 품종에서 발현되었다. HSPs는 토종수수 (TJS)에서만 발현 되는 것을 확인하였다. 이들 동정된 단백질들은 수수의 품종 별 특성을 이해하는데 중요한 단서를 제공할 것으로 예측된다.

• 한국안광학회지
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• 제12권1호
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• pp.9-15
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• 2007

본 연구는 UV-A 노출에 의해 유발된 안구 내에 존재하는 단백질 효소 중의 하나인 ribonuclease A(RNase A)의 변성을 차단할 수 있는 안경렌즈의 적절한 UV-A 차단율을 알아보기 위해 수행하였다. RNase A를 1, 3, 6, 24, 48, 72, 96시간 동안 365 nm의 UV-A에 노출시켜 노출시간에 따른 단백질의 변성 정도를 아크릴 아미드 겔 전기영동법으로 확인하였다. 또한, 20%, 50%, 80%, 99% UV 차단 효과를 가진 안경렌즈로 UV-A를 차단하였을 때 RNase A의 변성이 억제될 수 있는 지를 알아보았다. RNase의 변성은 1시간 동안의 UV 노출에 의해서도 유발되었으며, UV-A 노출시간이 길어질수록 그 정도는 심해졌다. 1시간의 UV-A 노출에 의해 유발된 미세한 RNase A의 변성은 20% 정도의 UV-A 차단으로 충분히 예방될 수 있었다. UV-A를 3시간 동안 노출하였을 때는 50%이상의 UV-A 차단율을 가진 렌즈가 RNase A의 변성을 막을 수 있었다. 6시간 동안 UV-A에 노출되었을 때에는 99%의 렌즈로 차단하였을 때조차도 RNase A의 변성이 완벽하게 차단되지 못했다. 그러나 96시간 동안 UV-A에 노출되었을 때 나타나는 심각한 단백질의 변성이 99%의 UV-A 차단 렌즈를 사용하였을 경우 크게 감소하였다.

• Journal of Pharmaceutical Investigation
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• 제34권5호
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• pp.369-377
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• 2004

The objective of this study was to investigate the patterns of protein leaching to an external phase during an ethyl acetate-based, double emulsion microencapsulation process. An aqueous protein solution (lactoglobulin, lysozyme, or ribonuclease; $W_1$) was emulsified in ethyl acetate containing poly-d,l-lactide-co-glycolide 75:25. The $W_1/O$ emulsion was transferred to a 0.5% polyvinyl alcohol solution saturated with ethyl acetate $(W_2)$. After the double emulsion was stirred for 5, 15, 30, or 45 min, additional 0.5% polyvinyl alcohol $(W_3)$ was quickly added into the emulsion. This so-called quenching step helped convert emulsion microdroplets into microspheres. After 2-hr stirring, microspheres were collected and dried. The degree of protein leaching to $W_2$ and/or $W_3$ phase was monitored during the microencapsulation process. In a separate, comparative experiment, the profile of protein leaching to an external phase was investigated during the conventional methylene chloride-based microencapsulation process. When ethyl acetate was used as a dispersed solvent, proteins continued diffusing to the $W_2$ phase, as stirring went on. Therefore, the timing of ethyl acetate quenching played an important role in determining the degree of protein microencapsulation efficiency. For example, when quenching was peformed after 5-min stirring of the primary $W_1/O$ emulsion, the encapsulation efficiencies of lactoglobulin and ribonuclease were $55.1{\pm}4.2\;and\;45.3{\pm}7.6%$, respectively. In contrast, when quenching was carried out in 45 min, their respective encapsulation efficiencies were $39.6{\pm}3.2\;and\;29.9{\pm}11.2%$. By sharp contrast, different results were attained with the methylene-chloride based process: up to 2 hr-stirring of the primary and double emulsions, less than 5% of a protein appeared in $W_2$. Afterwards, it started to partition from $W_1\;to\;W_2/W_3$, and such a tendency was affected by the amount of PLGA75:25 used to make microspheres. Different solvent properties (e.g., water miscibility) and their effect on microsphere hardening were to be held answerable for such marked differences observed with the two microencapsulation processes.

• 미생물학회지
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• 제52권3호
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• pp.243-248
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• 2016

RNase E는 대장균(Escherichia coli)에서 수많은 RNA의 가공 및 분해에 관여하는 필수적인 효소이다. RNase E의 효소 활성은 RraA와 RraB에 의해 조절된다. 그람양성균인 Streptomyces coelicolor는 RNase ES, RraAS1, RraAS2라고 명명되는 RNase E와 RraA의 동족체를 가지고 있다. 이 연구에서는 S. coelicolor 유래의 RraAS1이 E. coli에서 RNase E의 효소활성을 저해하는지 연구하였다. 대장균에서 RraAS1의 발현은 RNase E의 과발현에 의해 감소된 세포생장을 더욱 저하시켰으며, RNase E의 기질인 rpsO, ftsZ, rnhB mRNA의 양을 감소시키는 것을 확인 하였다. 이러한 RraAS1의 효과는 공동면역침전실험을 수행한 결과에서 유추할 수 있듯이, Rne 단백질과 RraAS1의 결합으로 유도되는 것으로 보인다. 이러한 결과는 RraAS1이 대장균에서 RNase E의 리보핵산 가수분해 활성을 유도함을 시사한다.

• 식물조직배양학회지
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• 제21권5호
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• pp.253-275
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• 1994

Many flowering plants possess genetically controlled self -incompatibility (SI) system that prevents inbreeding and promotes outcrosses. SI is usually controlled by a single, multiallelic S-locus. In gametophytically controlled system, SI results when the S-allele of the pollen is matched by one of the two S-alleles in the style, while in the sporophytic system self-incompatible reaction occurs by the interaction between the pistil genotype and genotype of, not the pollen, but the pollen parent In the former system the self-incompatible phenotype of pollen is determined by the haploid genome of the pollen itself but in the latter the pollen phenotype is governed by the genotype of the pollen parent along with the occurrence of either to-dominant or dominant/recessive allelic interactions. In the sporophytic type the inhibition reaction occurs within minutes following pollen-stigma contact, the incompatible pollen grains usually failing to germinate, whereas in gametophytic system pollen tube inhibition takes place during growth in the transmitting tissue of the style. Recognition and rejection of self pollen are the result of interaction between the S-locus protein in the pistil and the pollen protein. In the gametophytic SI the S-associated glycoprotein which is similar to the fungal ribonuclease in structure and function are localized at the intercellular matrix in the transmitting tissue of the style, with the highest concentration in the collar of the stigma, while in the sporophytic SI deposit of abundant S-locus specific glycoprotein (SLSG).is detected in the cell wall of stigmatic papillae of the open flowers. In the gametophytic system S-gene is expressed mostly at the stigmatic collar the upper third of the style length and in the pollen after meiosis. On the other hand, in the sporophytic SI S-glycoprotein gene is expressed in the papillar cells of the stigma as well as in e sporophytic tape is cells of anther wall. Recognition and rejection of self pollen in the gametophytic type is the reaction between the ribonuclease in the transmitting tissue of the style and the protein in the cytoplasm of pollen tube, whereas in the sporophytic system the inhibition of selfed pollen is caused by the interaction between the Sycoprotein in the wall of stigmatic papillar cell and the tapetum-origin protein deposited on the outer wall of the pollen grain. The claim that the S-allele-associated proteins are involved in recognition and rejection of self pollen has been made merely based on indirect evidence. Recently it has been verified that inhibition of synthesis of S$_3$ protein in Petunia inflata plants of S$_2$S$_3$ genotype by the antisense S$_3$ gene resulted in failure of the transgenic plant to reject S$_3$ pollen and that expression of the transgenic encoding S$_3$ protein in the S$_1$S$_2$ genotype confers on the transgenic plant the ability to reject S$_3$ pollen. These finding Provide direct evidence that S-proteins control the s elf-incompatibility behavior of the pistil.

• BMB Reports
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• 제42권12호
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• pp.829-833
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• 2009

Angiogenin is a member of the ribonuclease superfamily that induces the formation of new blood vessels. It has been suggested that the surface loop of angiogenin defined by residues 59-71 plays a special role in angiogenic function (1); however, the mechanism of action is not clearly defined. To elucidate the role of the surface loop on the structure, function and stability of angiogenin, three surface loop mutants were produced in which 14 amino acids in the surface loop of RNase A were substituted for the 13 amino acids in the corresponding loop of angiogenin. The structure, stability and biological functions of the mutants were then investigated using biophysical and biological approaches. Even though the substitutions did not influence the overall structure of angiogenin, they affected the stability and angiogenic function of angiogenin, indicating that the surface loop of angiogenin plays a significant role in maintaining the stability and angiogenic function of angiogenin.

• Applied Biological Chemistry
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• 제15권3호
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• pp.187-192
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• 1972

벼가 유아기에 냉해$(15^{\circ})C$로 인하여 생장이 정지될 때 RNA와 RNase활성도를 비교 연구하여 아래와 같은 결과를 얻었다. 1. 냉해 초기에는 RNA 함량과 RNase 활성도가 동시에 증가하고 3일 후에는 감소하였으나 5일 후에는 RNA 함량은 계속 감소하는 반면 RNase 활성도는 다시 증가 하였다. 2. 이러한 RNA의 변화는 ribosomal RNA의 변화에 기인함을 확인하였다. 3. RNase는 RNA의 합성 기능도 가지고 있는 것으로 사료되며 이 기능은 soluble RNA보다 ribosomal RNA와 관계가 있는 것으로 추측된다.

• Applied Biological Chemistry
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• 제15권3호
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• pp.181-186
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• 1972

소맥 자엽초를 일정한 길이로 절단하여 식물생장 조질제인 교dole acetic acid (IAA)와 abscieic acid (ABA)를 처리하여 아래와 같은 결과를 얻었다. 1. IAA는 고분자 핵산의 F2와 F3을 감소시킨 반면에 ABA는 F4를 증가 시켰다. 2. IAA는 G2 isozyme의 활성도를 증가시켰으나 G3 isogyme의 활성도는 ABA에 의하여 감소되었다. 3. 상기의 결과로 부터 IAA와 ABA는 핵산과 RNase에 대하여 어느정도 공통되는 효과를 갖는다는 것이 암시됨을 알수 있었다. 4. SH group을 저해하는 pb, p-hydroxymercury benfoa 둥에 의해서 발현되는 latent RNase의 활성도는 나타나지 않았다.

• Asian-Australasian Journal of Animal Sciences
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• 제3권2호
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• pp.115-120
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• 1990

Extracellular nuclease(s) in buffalo rumen fluid were purified from strained rumen fluid by a procedure involving Seitz filtration, acetone fractionation and gel filtration on Sephadex G-100. The enzyme resolved into two peaks exhibiting both DNase and RNase activities. The molecular weight of enzyme corresponding to peaks I and II were approximately 30,000 and 12,000 respectively. The properties of enzymes from the two peaks, however, were same. Optimum temperature for both DNase and RNase activities was at $50^{\circ}C$. Whereas DNase activity was stable upto $60^{\circ}C$, RNase activity was stable only up to $50^{\circ}C$. DNase activity recorded two pH optima, one at pH 5.5 and the other at pH 7.0. RNase activity recorded a broad pH optimum between pH 6.0-8.0. pH stability of the enzyme coincided with pH optima for both the activities. DNase activity was stimulated by $Mg^{2+}$ and $Mn^{2+}$ and inhibited by $Fe^{2+}$, $Zn^{2+}$, $Hg^{2+}$ and $Ag^+$. RNase activity was also stimulated by $Mg^{2+}$ and $Mn^{2+}$ and inhibited by $Cu^{2+}$, $Fe^{2+}$, $Zn^{2+}$, $Hg^{2+}$ and $Ag^+$. Reducing agents stimulated both the activities.