• 생명과학회지
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• 제27권12호
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• pp.1393-1402
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• 2017

식물에서, 칼슘-의존적 단백질 카이네즈(CDPKs)는 $Ca^{2+}$ 신호전달에서 중요한 $Ca^{2+}$ 수용체이다. 벼(Oryza sativa L.)의 CDPKs인 3개의 OsCPKs는 생물정보에 대한 분석이 이루어졌으나, OsCPK11 유전자는 연구가 완전히 수행되지 않았다. 다양한 조직에서 OsCPK11 유전자가 전사수준에서 발현한다는 것은 알려져 있으나, 단백질 수준에서 발현과 생화학적인 특징은 잘 알려져 있지 않다. 이 연구는 OsCPK11의 몇 가지 생화학적 특징을 알아보기 위해 이루어졌다. 먼저 in vitro에서 E. coli를 이용하여 GST-OsCPK11를 발현시키고, 카이네즈 활성 측정과 칼슘-의존적 단백질 카이네즈로서 OsCPK11의 생화학적 분석도 수행하였다. OsCPK11은 스스로 자가인산화하며, $Ca^{2+}$의 존재 하에서 기질로서 histone III-s와 MBP로 인산기 전달 작용을 수행한다. 재조합 OsCPK11의 활성은 $Mg^{2+}$에 의해 영향을 받으며, pH 7.0-7.5에서 최적의 활성을 보인다. 또한 OsCPK11의 활성은 높은 수준의 $Ca^{2+}$가 존재하는 조건에서는 $Mg^{2+}$, $Mn^{2+}$, $Na^+$의 영향을 받지 않는다. 또한 OsCPK11의 자가인산화는 OsCPK11의 $Ca^{2+}$ 민감도를 감소시키는 것으로 밝혀졌다. 마지막으로, OsCPK11의 N-말단 다양화 지역으로 토끼 항체를 만들었고, immunoblot을 기초로 polyclonal antibody는 95.5 kD의 GST-OsCPK11를 인식하는 것으로 나타났다. 이 결과는 벼의 $Ca^{2+}$ 매개 신호전달에서 OsCPK11의 기능을 더 잘 이해하는데 도움을 줄 것이며, 심화 연구를 위해 다양한 OsCPKs의 단백질 정보를 결정하는 것이 필요할 것이다.

• 한국임상수의학회지
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• 제18권4호
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• pp.390-396
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• 2001

CKP isoenzymes have a high level of efficaciousness as diagnostic and prognostic aids in various diseases. There is not any report on the total activity of CPK of non-human primates, let alone CPK isoenzyme patterns, in Korea. In this study, total activities and isoenzyme patterns of CPK were measured to obtain their reference values in domestically reared common marmosets, crab-eating macaques and Japanese macaques. We observed remarkable different values of serum total CPK from the primates used in this experiment. Serum CPK activities of Japanese macaques and crab-eating macaques were 275.8$\pm$158.1 IU/l and 396.7$\pm$697.4 IU/l, respectively, whereas those of common marmosets showed much higher value of 618.8$\pm$1,117.6 IU/l. In all common marmosets and crab-eating macaques, only CPK$_3$ ws observed. In five out of eight Japanese macaques, CPK$_3$ was the sole fraction but two animals showed CPK$_1$ and CPK$_3$ isoenzymes, and the remaining one had CPK$_2$ and CPK$_3$ fractions. There were some discrepancies in the pattern and ratio of isoenzyme fractions in Japanese macaques. In conclusion, values such as CPK and CPK isoenzyme patterns of investigated for the first time form non-human primates reared in Korea, could be reference values for the optimal diagnosis and therapy diseases of the corresponding animal species.

• 한국임상수의학회지
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• 제9권2호
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• pp.417-426
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• 1992

Total CPK activities and CPK isoenzymes fractions of the sera and some organ tissues of dogs were examined to acquire the basic data of canine CPK available in clinical practice. In addition fluctuation of serum total CPK activities and CPK isoenzymes fractions is artificially induced canine myocardial infarctino were investigated to know the availabity of them as indicators for the diagnosis of myocardial infarction. For the determination of serum total CPK activities, total 22 clinically healthy dogs(7 to 30 months old, 15 of female and 7 of male) were used and 15 out of 22 dogs were used for the determination of serum CPK isoenzymes fractions. For the determination of total CPK activities and CPK isoenzymes fractions. some organ tissues (the hearts, skeletal muscles and brains )from 3 dogs were examined. For the fluctuation of total CPK activities and CPK isoenzymes fractions in the sera from artificially induced canine acute myocardial infarction, 3 dogs of coronary artery ligated experimental group and 3 of control group were used. The results obtained were as follows ; 1. Serum total CPK activities of normal dogs were 106.2${\pm}$29.9(31.3∼148.1)IU/$\ell$. 2. The pattern of serum CPK isoenzymes fractions in normal dogs was high with decreasing order of CK$_1$>CK$_3$>CK$_2$. 3. Total CPK activities of organ tissues were high with decreasing order of the skeletal muscles > the hearts > the brains. 4. The pattern of CPK isoenzymes fractions of the organ tissues was high with decreasing order of CK$_3$>CK$_2$ in the hearts and only CK$_3$(100%) was detected in the skeletal muscles. Further they were high with decreasing order of CK$_1$>CK$_3$>CK$_2$ in the trains. 5. Serum total CPK activities in experimental group were changed with higher values than those of control group. 6. In the fluctuation of serum CPK isoenzymes fractions the CK$_1$ CK$_2$ and CK$_3$ values were changed with higher values than those of control group. 7. It was become clear that the finding of Increase of serum total CPK activities, and CK$_2$ and CK$_3$ was important for the diagnosis of myocardial infarction.

• 생명과학회지
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• 제31권2호
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• pp.115-125
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• 2021

칼슘-의존성 단백질 카이네이즈(CDPK)는 식물의 Ca2+ 매개 신호 전달에 필수적인 역할을 한다. CDPK는 염분, 저온, 가뭄 등과 같은 비생물적 스트레스에 대한 식물의 반응을 조절하는 데 관여하는 것으로 알려져 있다. 벼의 CDPK는 31개의 유전자로 구성된 거대 다유전자군으로 되어 있지만, 단지 일부 벼의 CDPK 기능만이 확인되었다. 따라서, 벼에서 OsCPK11의 기능을 알아보기 위해, 이 연구는 염분, 저온 및 가뭄과 같은 비생물적 스트레스 조건에서 벼의 야생형과 ND0001 oscpk11 돌연변이체의 OsCPK11 발현 분석에 초점을 맞추었다. 염분, 저온, 가뭄 스트레스 처치를 위해 유식물을 각각 200 mM NaCl, 4℃, 20% PEG 6,000에 노출시켰다. 야생형과 ND0001 돌연변이체에서 OsCPK11의 발현을 확인하기 위해 RT-PCR과 quantitative real-time PCR을 수행하였다. RT-PCR 결과에 의하면, 야생형과 이형접합성 돌연변이체에서는 OsCPK11 전사체가 검출되었지만, 동형접합성 돌연변이체에서는 검출되지 않았다. Quantitative real-time PCR 결과에 의하면 야생형에서 염분, 저온, 가뭄 스트레스에 의해 OsCPK11의 상대적인 발현이 증가하였으며, 각각 24시간, 6시간, 24시간 후 최대 수준에 도달하였다. ND0001 동형접합성 돌연변이체의 OsCPK11의 상대적 발현은 야생형에 비해 현저히 감소하였다. 이러한 결과는 oscpk11 동형접합성 돌연변이체에서는 OsCPK11발현을 완전히 저해하며, OsCPK11유전자 발현 조절이 염분, 저온 및 가뭄 스트레스 신호 전달 과정에 관여할 수 있음을 의미한다.

• 한국임상수의학회지
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• 제9권2호
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• pp.433-449
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• 1992

Total CPK activities and CPK isoenzymes fractions of the sera and tissues were examined to obtain the physiological basic data of ruminant available in veterinary clinical practice. For the sera total CPK activities and CPK isoenzymes fractions, total 39 clinically healthy Korean native goats (3 to 10 months old, IS of female and 18 of male) and 6 of Korean native goats (1 to 2 years old, 3 of female and 3 of male) were used. Seventeen Korean native cattle (3 to 6 years old, 10 of female and 7 of male) and 27 Holstein-Friesian cattle (2 to 8 months old, 7 of female and 3 to 12 years old, 20 of female) were also examined for the sera total CPK activities and CPK isoenzymes fractions. For the total CPK activities and CPK isoenzyme fractions, 3 of female Korean native goats (7 months old), 3 of female Korean native cattle (2 years old) and 3 of dairy cattle (2 years old, 2 of female and 1 of male) were used. The tissues examined were the cerebrum (2 of Korean native cattle), spinal cord (1 of Korean native cattle), heart, lung, diaphragm, reticulum, liver, spleen, kidney, jejunum. colon and femoral muscle. The results obtained were as follows : 1. In Korean native goats less than 1-year-old. serum total CPK activities were 67.8${\pm}$17.7(39.0~96.5) IU/$\ell$ in female and 63.4${\pm}$19.0(28.7~94.4) IU/$\ell$ in male. Further they were 67.0${\pm}$5.3(59.5~70.7) IU/$\ell$ and 54.5${\pm}$11.1(39.1~69.4) IU/$\ell$ in female and male Korean native goats over 1-year-old, respectively. Serum total CPK activities of female were slightly higher than those of male. Significance between age and sex was not found. 2. Serum total CPK activities were 56.8${\pm}$19.7(27.6~90.5) IU/$\ell$ and 65.6${\pm}$10.8(52.8~78.0) IU/$\ell$ in female and male Korean native adult cattle, respectively, Serum total CPK activities of male were slightly higher than those of female, but they were not significant 3. Serum total CPK activities we,e 72.5${\pm}$8.2(57.2~83.2) IU/$\ell$ and 60.8${\pm}$12.5(42.7~80.6) IU/$\ell$ in calves and adult of dairy acttle, respectively. Serum total CPK activities of calves were significantly higher than those of adult(p<0.05). 4. In Korean native goats less than 1-year-old, serum CPK isoenzymes fractions were high with decreasing order of MM>MB>BB and MM>BB>MB in female and male, respectively. Further they were high with decreasing order of MM>MB>BB and MM>B8>MB in female and male Korean native goats over 1-year-old, respectively. The main fractions of CPK isoenzymes were MM in sera of Korean native goats. 5. Serum CPK Isoenzyme fractions were high with decreasing order of MM>MB>BB In both female and male of Korean native cattle. The main fraction among them was MM. 6. Serum CPK isoenzymes fractions were high with decreasing order of MM>BB>MB in both calves and adult of dairy cattle. The main fraction among them was MM. 7. Total CPK activities were high with decreasing order of the femoral muscle>kidney>reticulum>diaphragm>liver>spleen>heart>colon>lung>jejunum in Korean native goats. 8. Total CPK activities were high with decreasing order of the spinal cord >cerebrum>femoral muscle>reticulum>kidney>liver>spleen>diaphragm>lung>colon>heart>jejunum in Korean native cattle. 9. Total CPK activities were high with decreasing order. of the femoral muscle >liver>retoculum>kidney>heart>colon>lung>spleen>jejunum>diaphrasm in dairy cattle. 10. The pattern of the cardiac CPK isoenzymes fractions was identical in Korean native goats, Korean native cattle and dairy cattle. They were high in the order of MM>MB without BB fractions and the main fraction was MM. 11. The pattern of the pulmonary CPK isoenzymes fractions was the same Korean native goats, Korean native cattle and dairy cattle. They were high with decreasing order of MM>MB>BB and the main fraction among them was MM. 12. The pattern of CPK isoenzymes fractions of the diaphragm was Identical in Korean native goats and Korean native cattle. They were high with decreasing order of MM >BB >MB except dairy cattle (MM>MB>BB) but the main fraction among them was MM. 13. The pattern of the reticular CPK isoenzymes fractions was identical in Korean native cattle and dairy cattle. They were high with decreasing order of BB >MM >MB except Korean native goats(BB>MB>MM) but the main, fraction among them was BB 14. The pattern of the hepatic CPK isoenzymrs fractions was identical in Korean native cattle and dairy cattle. They were high with decreasing order of MB >BB >MM except Korean native goats(MB>MM>BB)but the main fraction was MB. 15. The splenic CPK isoenzymes fractions showed different pattern. They were high with decreasing order of MB>BB>MM, MM>BB>MB and BB>MB>MM in Korean native goats, Korean native cattle and dairy cattle, respectively. The main fraction among them was different from each other. 16. The pattern of the renal CPK isoenzymes fractions was identical in Korean native cattle and dairy cattle. They were high with decreasing order of MM >MB>BB except Korean native goats(BB>MB>MM). 17. The CPK isoenzymes fractions of the Jejunums showed different pattern. They were high with decreasing order MM>MB>BB, MM>BB>MB and BB>MM>MB in Korean native goats, Korean native cattle and dairy cattle, respectively. The main fractions were MM In Korean native goats and Korean native cattle, and BB in dairy cattle. 18. The colonic CPK isoenzymes fractions showed different pattern. They were high with decreasing order of MM>MB>BB, MM>BB>MB and BB>rrfB>MM in Korean native goats, Korean native cattle and dairy cattle, respectively. The main fractions were MM in Korean native goats and Korean . native cattle, and BB in dairy cattle. 19. The cerebral CPK isoenzymes fractions were high with decreasing order of BB >MM without MB detected in Korean native cattle and those of spinal cord were high with decreasing order of BB >MM >MB. The main fractions in both cerebrum and spinal cord were BB.

• 생명과학회지
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• 제27권11호
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• pp.1233-1244
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• 2017

Calcium-dependent protein kinases (CDPKs)는 칼슘 이온을 매개로 한 신호전달 경로에서 중요한 역할을 한다. 벼(Oryza sativa)에는 29개의 CDPKs가 확인되었지만 그들의 기능은 완벽히 밝혀지지 않았다. 이 연구는 OsCPK11 유전자에 초점을 맞춰 그것의 기능적인 특징을 조사하였다. 벼의 어린 잎, 성장한 잎, 꽃에서 OsCPK11 유전자의 조직-특이적 발현이 확인되었고, 지베렐린이 처리된 벼의 호분층에서도 이 유전자의 발현을 확인할 수 있었다. Tos-17이 삽입된 oscpk11의 표현형에서 돌연변이체 각각의 키는 야생형과 구분되지 않았지만, 영과의 수나 무게는 통계적으로 유의미한 차이가 있었다. 덧붙여 많은 돌연변이체 낟알의 배젖에서 white belly materials이 확인되었다. OsCPK11의 cDNA가 cloning되었고, 약 60.5 kD인 OsCPK11 단백질이 GST affinity chromatography와 SDS-PAGE에 의해 얻어졌다. 아미노산 서열 분석을 통해 OsCPK11이 전형적인 CDPKs의 구조적 특징을 가짐을 알 수 있었다. 이 결과는 OsCPK11유전자의 기능과 식물에서 칼슘 이온을 매개로 한 신호전달 경로에 CDPK 역할의 유용한 정보를 제공해줄 것이다.

• BMB Reports
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• 제39권5호
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• pp.607-617
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• 2006

A novel calcium-dependent protein kinase gene (designated as IiCPK2) was cloned from tetraploid Isatis indigotica. The full-length cDNA of IiCPK2 was 2585 bp long with an open reading frame (ORF) of 1878 bp encoding a polypeptide of 625 amino acid residues. The predicted IiCPK2 polypeptide included three domains: a kinase domain, a junction domain (or autoinhibitory region), and a C-terminal calmodulin-like domain (or calcium-binding domain), which presented a typical structure of plant CDPKs. Further analysis of IiCPK2 genomic DNA revealed that it contained 7 exons, 6 introns and the length of most exons was highly conserved. Semi-quantitative RT-PCR revealed that the expression of IiCPK2 in root, stem and leaf were much higher in tetraploid sample than that in diploid progenitor. Further expression analysis revealed that gibberellin ($GA_3$), NaCl and cold treatments could up-regulate the IiCPK2 transcription. All our findings suggest that IiCPK2 might participate in the cold, high salinity and GA3 responsive pathways.

• The Journal of Korean Physical Therapy
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• 제11권1호
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• pp.103-110
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• 1999

The purpose of this study was to analyze the correlation coefficients between delayed muscle soreness after eccentric exercise, muscle strength, CPK, and ALD. Subjects of this study were 9 male students. CPK, ALD that known as the indicators of muscle damage and eccentric strength of the Lt elbow flexors were measured prior to exercise. After measurements Hey Perform an isokinetic eccentric exercise of flexor group of 1 elbow(10 repetitions 3 bouts) by make use of KIN-COM isokinetic device. After exercise, CPK, ALD, and peak torque of elbow flexor group was measured at the just after exercise, 24hr after, 72hr after. Also muscle soreness level was evaluated at same intervals by make use of VAS(visual analog scale). The results were as follows : 1 . There was significant differencess or muscle soreness, CPK, ALD, peak torque with the passage or recovery time (p<0.001). 2. There was negative correlation coefficients between CPK and peak torque at 24hr after(p<0.05). 3. There was positive correlation coefficients between ALD and CPK at 24hr after(p<0.05). 4. There was no correlation coefficients between other variables but ALD was the most important factors that can be explains the muscle sur eness very well.

• 한국동물위생학회지
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• 제17권2호
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• pp.114-121
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• 1994

Clinically healthy 12 female Manina breed chicken (6 of 75 days old : group 46 of 145 days old： group B, female) were examined to establish physiological basic data on organ tissues total Creatine phosphokinase (CPK) activities and CPK isoenzymes fractions. The tissues examined were the Lung, Heart, Liver, Proventriculus, Gizzard, Duodenum, Colon and Muscle. The results obtained were summarized as follows : 1. Total CPK activities were high with decreasing order of the Muscle > Proventriculus >Giz-zard >Heart >Duodenum> Colon >Lung >Liver in group A and Muscle >Proventriculus >Giz-zard >Heart > Colon >Duodenum > Lung > Liver in group B. Significance of total CPK activities in group difference was only found Colon, group B showed higher values than that of group A (p< 0.01). 2. In the pattern of CPK isoenzymes fractions, Lung, Heart, Liver, Proventriculus, Gizzard, Duodenum and Muscle were high with decreasing order of CK2 >CK3, Colon showed the pattern with decreasing order of CK3 >CK2. Significance of CPK isoenzymes fractions in group difference was only found Liver, CK2 in group B(P<0.01) and CK3 in group A(P<0.01) were higher than that of the other group.

• 생명과학회지
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• 제30권2호
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• pp.137-146
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• 2020

식물에서 Ca²⁺는 세포의 중요한 2차 신호 전달 분자 중 하나이다. Ca²⁺ 및 인산화 효소의 센서 단백질인 칼슘-의존성 단백질 카이네즈(CDPKs)는 식물 세포에서 가장 풍부한 세린/트레오닌 키나아제이다. 이들은 다양한 자극에 대한 신호를 변환하여 식물에서 특정 반응을 일으킨다. 벼에는 31개의 CDPK 유전자 족이 확인되었다. 그들은 주로 식물의 생장과 발달에 관여하며 다양한 스트레스 조건에 반응하여 기능을 하는 것으로 알려져 있다. 그러나 CDPK 단백질의 생화학적 특성에 대해서는 알려진 바가 별로 없다. 이 연구에서는 벼의 CDPK 중 하나인 OsCPK11을 부분 정제하여 그 생화학적 특성을 조사하고자 하였다. 벼 유식물에서 3단계 칼럼 크로마토그래피 과정을 거쳐 부분 정제된 OsCPK11을 얻었다. 정제 과정에는 DEAE를 사용한 음이온 교환 크로마토그래피, Phenyl-Sepharose를 사용한 소수성 상호작용 크로마토그래피 및 Sephacryl-200HR를 사용한 겔 여과 크로마토그래피를 포함하였다. 부분 정제된 OsCPK11은 분자량이 54kDa이며 소수성 수지와 강한 소수성 상호작용을 보였다. 부분 정제된 OsCPK11으로 in vitro kinase assay를 실시한 결과, OsCPK11은 Ca²⁺-의존성 자가인산화 활성을 가짐을 보여 주었다. OsCPK11은 histone III-S를 인산화 하였으며, 카이네즈 활성의 최적 pH는 7.5-8.0이었다. Native OsCPK11은 이전에 연구된 재조합 OsCPK11과 몇 가지 생화학적 특징을 공유하였는데, 둘 다 Ca²⁺-의존성 자가인산화 활성을 나타냈다. 또한, 둘 모두 카이네즈 활성을 위한 기질로서 histone III-S를 선호하였으며, Ca²⁺ 의존성을 보여 주었다.