• Journal of Life Science
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• v.27 no.12
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• pp.1393-1402
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• 2017

In plants, calcium ($Ca^{2+}$)-dependent protein kinases (CDPKs) are important sensors of $Ca^{2+}$ signals. Previous research demonstrated the expression of the OsCPK11 gene in various tissues at the transcription level, but its developmental and biochemical functions at the protein level were not determined. This study was aimed to identify biochemical characteristics of OsCPK11. GST- OsCPK11 was expressed in E. coli and used for an in vitro kinase assay. Biochemical analyses identified OsCPK11 as a CDPK. OsCPK11 autophosphorylated itself and transphosphorylated histone III-s and MBP as substrates in the presence of $Ca^{2+}$. The activity of the recombinant OsCPK11 was influenced by $Mg^{2+}$, with optimum activity detected at pH 7.0-7.5. OsCPK11 activity was not affected by $Mg^{2+}$, $Mn^{2+}$, or $Na^+$ in the presence of a high level of $Ca^{2+}$. Autophosphorylation of OsCPK11 decreased $Ca^{2+}$ sensitivity of OsCPK11. An anti-OsCPK11 rabbit antibody recognized 95.5 kD of GST-OsCPK11, as shown by an immunoblot analysis. These results shed light on the function of OsCPK11 in $Ca^{2+}$-mediated signaling in rice.

• Journal of Veterinary Clinics
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• v.18 no.4
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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.

• Journal of Veterinary Clinics
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• v.9 no.2
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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.

• Journal of Life Science
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• v.31 no.2
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• pp.115-125
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• 2021

Calcium-dependent protein kinases (CDPKs) are known to be involved in regulating plant responses to abiotic stresses such as salinity, cold temperature and dehydration,. Although CDPKs constitute a large multigene family consisting of 31 genes in rice, only a few rice CDPKs' functions have been identified. Therefore, in order to elucidate the functions of OsCPK11 in rice, this study was intended to focus on the expression pattern analysis of OsCPK11 in wild type and ND0001 oscpk11 mutant plants under these abiotic stresses. For the salt, cold and drought stress treatment, seedlings were exposed to 200 mM NaCl, 4℃ and 20% PEG 6,000, respectively. RT-PCR and quantitative real-time PCR were performed to determine the expression patterns of OsCPK11 in wild type and ND0001 mutant plants. RT-PCR results showed that OsCPK11 transcripts in the wild type and heterozygous mutant were detected, but not in the homozygous mutant. Real-time PCR results showed that relative expression of OsCPK11 of wild type plants was increased and reached to the highest level at 24 hr, at 6 hr and at 24 hr under salt, cold and drought stress conditions, respectively. Relative expression of OsCPK11 of ND0001 homozygous plant was significantly reduced compared to that of wild type. These results suggested that oscpk11 homozygous mutant knocks out OsCPK11 and OsCPK11 might be involved in salt, cold and drought stress signaling by regulating its gene expression.

• Journal of Veterinary Clinics
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• v.9 no.2
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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.

• Journal of Life Science
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• v.27 no.11
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• pp.1233-1244
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• 2017

CDPKs have pivotal roles in plant $Ca^{2+}$-mediated transduction signaling. A total of 29 CDPK genes have been identified in rice (Oryza sativa L.), but their key functions have not been completely noted. This study focused on the OsCPK11 gene, which has not been studied, to determine its functional characteristics. A study of tissue-specific expressions revealed that the OsCPK11 gene is expressed in young leaves, mature leaves and flowers of rice. An expression of the gene was also confirmed in gibberellin-treated aleurone layers of rice. Regarding the phenotypic characteristics of Tos17-inserted OsCPK11 mutants, the heights of the mutants were not distinguishable from the heights of wild type plants, but the number of caryopses and the caryopses' weights were significantly statistically different. In addition, many grains of the mutants had white belly materials in their endosperm. The cDNA of the OsCPK11 was cloned, and an OsCPK11 protein of about 60.5 kD was obtained by using a GST affinity chromatography and an SDS-PAGE. An analysis of the amino-acid sequence of the protein indicated that the OsCPK11 protein has the structural characteristics of typical CDPKs. The results provided useful information about the functions of the OsCPK11 gene and further noted the roles CDPKs have in $Ca^{2+}$-mediated signaling in plants.

• BMB Reports
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• v.39 no.5
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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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• v.11 no.1
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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.

• Korean Journal of Veterinary Service
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• v.17 no.2
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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.

• Journal of Life Science
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• v.30 no.2
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• pp.137-146
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• 2020

Calcium is one of the important secondary signaling molecules in plant cells. Calcium-dependent protein kinases (CDPK)-the sensor proteins of Ca²⁺ and phosphorylating enzymes-are the most abundant serine/threonine kinases in plant cells. They convert and transmit signals in response to various stimuli, resulting in specific responses in plants. In rice, 31 CDPK gene families have been identified, which are mainly involved in plant growth and development and are known to play roles in response to various stress conditions. However, little is known about the biochemical characteristics of CDPK proteins. In this study, OsCPK11-a CDPK in rice-was partially purified, and its biochemical characteristics were found. Partially purified OsCPK11 from rice seedlings was obtained by three-step column chromatography that involved anion exchange chromatography consisting of DEAE, hydrophobic interaction chromatography consisting of phenyl-Sepharose, and gel filtration chromatography consisting of Sephacryl-200HR. An in vitro kinase assay using partially purified OsCPK11 was also performed. This partially purified OsCPK11 had a molecular weight of 54 kDa and showed a strong hydrophobic interaction with the hydrophobic resin. In vitro kinase assay showed that the OsCPK11 also had Ca²⁺-dependent autophosphorylation activity. The OsCPK11 phosphorylated histone III-S, and the optimum pH for its kinase activity was found to be 7.5~8.0. The native OsCPK11 shared several biochemical characteristics with recombinant OsCPK11 studied previously, and both had Ca²⁺-dependent autophosphorylation activity and favored histone III-S as a substrate for kinase activity, which also had a Ca²⁺-dependence.