• 한국동물학회지
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• 제36권3호
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• pp.347-352
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• 1993

N-ethylmaleimide는 낮은 농도에서 Myosin heavy chain의 활성화 부위에 존재하는 2개의 황화기에 선택적으로 결합하는 것으로 알려져 있다. 계 근조직에서 분리된 $Ca^2$+-의존성 단백질 분해효소, Calpain은 이와같이 알킬화된 Myosin heavy chain을 알킬화 되지 않은 것에 비하여 우선적으로 분해하는 것으로 나타났다. 또한, 황화기를 특이하게 산하시키는 KMnO$_4$가 처리된 Myosin heavy chain도 산화되지 않은 것에 비하여 훨씬 빠른 속도로 분해됨을 관찰하였다. 뿐만아니라, N-ethylmaleimide나 KMnO$_4$의 처리는 농도-의존적으로 myosin에 의한 ATP 분해를 불활성화 시키었다. 이러한 결과는 활성화 부위에 존재하는 황화기의 화학적 변형은 Myosin hsavy chain이 Calapin과 같은 세포내 단백질 분해효소에 의하여 인식되는 기구임을 시사한다.

• 한국축산식품학회지
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• 제34권5호
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• pp.656-664
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• 2014

The purpose of this study was to analyze myosin heavy chain (MHC) isoforms in bovine longissimus thoracis (LT) muscle by liquid chromatography (LC) and mass spectrometry (MS). LT muscles taken from Hanwoo (Korean native cattle) steer (n=3) used to separate myosin bands by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The peptide queries were obtained from the myosin bands by LC-MS/MS analysis following in-gel digestion with trypsin. A total of 33 and 43 queries were identified as common and unique peptides, respectively, of MHC isoforms (individual ions scores >43 indicate identity or extensive homology, p<0.05). MHC-1 (IIx), -2 (IIa), -4 (IIb), and -7 (slow/I) were identified based on the Mowse score (5118, 3951, 2526, and 2541 for MHC-1, -2, -4, and -7, respectively). However, more analysis is needed to confirm the expression of MHC-4 in bovine LT muscle because any query identified as a unique peptide of MHC-4 was not found. The queries that were identified as unique peptides could be used as peptide markers to confirm MHC-1 (14 queries), -2 (8 queries), and -7 (21 queries) in bovine LT muscle; no query identified as a unique peptide of MHC-4 was found. LC-MS/MS analysis is a useful approach to study MHC isoforms at the protein level.

• Journal of Animal Science and Technology
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• 제65권3호
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• pp.511-518
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• 2023

This study examined the association between functional sequence variants (FSVs) of myosin heavy chain 3 (MYH3) genotypes and collagen content in a Landrace and Jeju native pig (JNP) crossbred population. Four muscles (Musculus longissimus dorsi, Musculus semimembranosus, Musculus triceps brachii, and Musculus biceps femoris) were used for the analysis of meat collagen content, and the same animals were genotyped for the FSVs of the MYH3 gene by using PCR-RFLP (polymerase chain reaction-restriction fragment length polymorphism). Three FSVs of MYH3 genotypes were identified and had genotype frequencies of 0.358, 0.551, and 0.091 for QQ, Qq, and qq, respectively. QQ animals for the FSVs of the MYH3 genotypes showed higher collagen content in their M. longissimus dorsi (p < 0.001), M. semimembranosus (p < 0.001), M. triceps brachii (p < 0.001), and M. biceps femoris (p < 0.001) than qq homozygous animals. After the validation of this result in other independent populations, the FSVs of MYH3 genotypes can be a valuable genetic marker for improving collagen content in porcine muscles and can also be applied to increase the amount of collagen for biomedical purposes.

• Asian-Australasian Journal of Animal Sciences
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• 제21권12호
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• pp.1703-1709
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• 2008

As genetic markers, single nucleotide polymorphisms (SNP) are very appropriate for the development of genetic tests for economic traits in livestock. Several microsatellite markers have been identified as useful markers for the genetic improvement of Hanwoo. Among those markers, ILSTS035 was recently mapped at a similar position with four SNPs (AH1_11, AH1_9, 31465_446, and 12273_165) in a linkage map of EST-based SNP in BAT6. Among the four SNPs, two SNPs (31465_446 and 12273_165) were analyzed using BLAST at the NCBI web site. The sequences including the 12273_165 SNP were identified at the intron region within the LOC534614 gene on the gene sequence map (Bos taurus NCBI Map view, build 3.1). The LOC534614 gene represents a protein similar to myosin heavy chain, fat skeletal muscle, embryonic isoform 1 in the dog, and myosin_1 (Myosin heavy chain D) in Macaca mulatta. In cattle, the myosin heavy chain was associated with muscle development. The phenotypic data for growth and carcass traits in the 415 animals were analyzed by the mixed ANCOVA (analysis of covariance) linear model using PROC GLM module in SAS v9.1. By the genotyping of Hanwoo individuals (n = 415) to evaluate the association of SNP with growth and carcass traits, it was shown that the 12273_165 SNP region within LOC534614 may be a candidate marker for growth. The results of the statistical analyses suggested that the genotype of the 12273_165 SNP significantly affected birth weight, weight of the cattle at 24 months of age, average daily gain and carcass cold weight (p<0.05). Consequently, the 12273_165 SNP polymorphisms at the LOC534614 gene may be associated with growth in Hanwoo, and functional validation of polymorphisms in LOC534614 should be performed in the future.

• 한국식품영양과학회지
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• 제33권10호
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• pp.1668-1675
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• 2004

근원섬유단백질과 알칼리 용액으로 처리하여 회수한 단백질의 가열 중 ANS 소수성의 변화, IR스펙트럼의 변화, SH기의 변화, 전기영동상의 변화 및 엔탈피의 변화로 비교하여 회수 단백질의 가열 변성기구를 조사하였다. 갈고등어와 냉동 꼬마민어의 근원섬유단백질은 가열 온도의 상승과 더불어 소수성 잔기가 외부로 노출되고 소수성 상호작용은 6$0^{\circ}C$ 부근에서 최대로 일어나지만, 근형질 단백질을 포함하는 회수단백질은 근원섬유 단백질과 ANS 소수성에서 차이를 보였다. pH가 증가함에 따라 1636 $cm^{-1}$ /에 해당하는 peak가 증가하였다. 반응성 SH기와 총 SH기 수의 차이는 pH 7.0과 pH 10에서 비교적 큰 것으로 나타났다. SDS-PAGE 상에서 pH가 상승함에 따라 myosin heavy chain의 중합체가 확인되었다. 알칼리 공정으로 제조한 회수 단백질을 시차주사열량계로 분석한 결과, 회수 단백질은 33.1,44.3 및 65.5$^{\circ}C$에서 전이 온도가 나타나며, myosin의 변성 에 해당하는 온도인 51.7$^{\circ}C$의 peak는 보이지 않았다. 회수 단백질의 가열 젤은 극단적 인 pH 처리에 의한 $\alpha$-helix 구조의 $\beta$-sheet 구조 전환, 가열에 의한 S-S 결합의 형성과, myosin heavy chain 중합체 형성에 의한 것으로 보인다.

• 한국수산과학회지
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• 제29권3호
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• pp.296-308
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• 1996

혈합육어 (멸치와 전어)와 백색육어 (농어와 도다리)의 육과 내장에서 추출한 조효소에 대하여 천연기질 및 합성 기질에 대한 효소 활성과 disc-PAG 전기영동에서 분리된 효소 단백질의 분포를 선택적 합성기질을 써서 비교 검토하였다. 육에는 SH-proteinase인 cathepsin L, B, 및 H 유사 효소가 분포하는 것을 확인 할 수 있었으며, 이 밖에도 cathepsin G 및 D도 다소 분포하는 것으로 추정되었다. 그리고 내장에는 강한 활성의 chymotrypsin 및 trypsin 유사 효소가 주로 분포하고 있음이 확인되었다. 육과 내장에서 추출한 조효소들은 공통적으로 멸치, 전어, 도다리 그리고 농어의 순으로 활성이 높은 것을 알 수 있었으며, 전체적으로 혈합육어에 분포되어 있는 효소가 백색육어류에 분포되어 있는 효소에 비하여 약 100배정도 높았다. 육과 내장의 조효소를 방어의 근원섬유단백질에 작용시킨 결과, 육 중의 조효소는 myosin heavy chain의 분해가 두드러졌으며, 특히 멸치의 효소가 근원섬유 단백질의 구성 subunit 모두에 대하여 강한 분해능을 보였다. 내장 조효소의 경우는 반응 초기에 myosin heavy chain과 actin을 현저하게 분해시켰으며, 혈합육어인 멸치와 전어 조효소의 분해능이 백색육어인 농어와 도다리의 조효소보다 강한 것을 확인 할 수 있었다. 육의 조효소는 어종에 따라 서로 다른 분해 양상을 보였으며, 내장 조효소는 혈합육어와 백색육어의 효소 활성뿐만 아니라 효소의 분포상으로도 많은 차이가 있었다.

• 한국동물위생학회지
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• 제31권4호
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• pp.449-456
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• 2008

The differentiation of mouse embryonic stem(ES) cell into smooth muscle cells(SMC) may play a major role in cardiovascular development and under pathophysiological conditions. Therefore, in the present study, we have examined the differentiation of ES cells and its related gene expression. SMC differentiation was indicated by cellular morphology and time-dependent induction of dibutyryl adenosine 3,5-cyclic monophosphate(DBcAMP)and retinoic acid(RA) on smooth muscle ${\alpha}$-actin($SM{\alpha}A$), smooth muscle myosin heavy chain(SMMHC) gene expression. The control was undifferentiated ES cells(protein expressions represent 50-60kDaOct-4). The results of this study show that morphology of embryoid body and confirmation of $SM{\alpha}A$ expression by immunocytochemistry. Moreover, SMMHC and desmin expression was significantly increased by time dependent manner(5, 7, 15 days), in contrast to $SM{\alpha}A$ expression was slightly decreased on 15days. In conclusion, DBcAMP and RA stimulate mouse ES cells differentiation into SMC and enhanced $SM{\alpha}A$, SMMHC and desmin expression.

• The Korean Journal of Physiology and Pharmacology
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• 제3권2호
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• pp.223-230
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• 1999

Alterations of cardiovascular function associated with various thyroid states have been studied. In hyperthyroidism left ventricular contractility and relaxation velocity were increased, whereas these parameters were decreased in hypothyroidism. The mechanisms for these changes have been suggested to include alterations in the expression and/or activity levels of various proteins; ${\alpha}-myosin$ heavy chain, ${\beta}-myosin$ heavy chain, ${\beta}-receptors,$ the guanine nucleotide-binding regulatory protein, and the sarcolemmal $Ca^{2+}-ATPase.$ All these cellular alterations may be associated with changes in the intracellular $Ca^{2+}$ concentration. The most important regulator of intracellular $Ca^{2+}$ concentration is the sarcoplasmic reticulum (SR), which serves as a $Ca^{2+}$ sink during relaxation and as a $Ca^{2+}$ source during contraction. The $Ca^{2+}-ATPase$ and phospholamban are the most important proteins in the SR membrane for muscle relaxation. The dephosphorylated phospholamban inhibits the SR $Ca^{2+}-ATPase$ through a direct interaction, and phosphorylation of phospholamban relieves the inhibition. In the present study, quantitative changes of $Ca^{2+}-ATPase$ and phospholamban expression and the functional consequences of these changes in various thyroid states were investigated. The effects of thyroid hormones on (1) SR $Ca^{2+}$ uptake, (2) phosphorylation levels of phospholamban, (3) SR $Ca^{2+}-ATPase$ and phospholamban protein levels, (4) phospholamban mRNA levels were examined. Our findings indicate that hyperthyroidism is associated with increases in $Ca^{2+}-ATPase$ and decreases in phospholamban levels whereas opposite changes in these proteins occur in hypothyroidism.

• 한국축산식품학회지
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• 제40권1호
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• pp.132-144
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• 2020

The aim of this study was to optimize staining procedures for muscle fiber typing efficiently and rapidly in bovine and porcine skeletal muscles, such as longissimus thoracis, psoas major, semimembranosus, and semitendinosus muscles. The commercially available monoclonal anti-myosin heavy chain (MHC) antibodies and fluorescent dye-conjugated secondary antibodies were applied to immunofluorescence histology. Two different procedures, such as cocktail and serial staining, were adopted to immunofluorescence analysis. In bovine muscles, three pure types (I, IIA, and IIX) and one hybrid type, IIA+IIX, were identified by the cocktail procedure with a combination of BA-F8, SC-71, BF-35, and 6H1 anti-MHC antibodies. Porcine muscle fibers were typed into four pure types (I, IIA, IIX, and IIB) and two hybrid types (IIA+IIX and IIX+IIB) by a serial procedure with a combination of BA-F8, SC-71, BF-35, and BF-F3. Unlike for bovine muscle, the cocktail procedure was not recommended in porcine muscle fiber typing because of the abnormal reactivity of SC-71 antibody under cocktail procedure. Within the four antibodies, combinations of two or more anti-MHC antibodies allowed us to distinguish pure fiber types or all fiber types including hybrid types. Application of other secondary antibodies conjugated with different fluorescent dyes allowed us to get improved image resolution that clearly distinguished hybrid fibers. Muscle fiber characteristics differed depending on species and muscle types.

• 한국진공학회:학술대회논문집
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• 한국진공학회 2015년도 제49회 하계 정기학술대회 초록집
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• pp.158-158
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• 2015

Myoblast are myogenic precursors that proliferate, activate, and differentiate on muscle injury to sustain the regenerative capacity of skeletal muscle; The neuronal isoform of nitric oxide synthase (nNOS, termed also NOS-I) is expressed in normal adult skeletal muscle, suggesting important functions for Nitric oxide (NO) in muscle biology1,2,3. However, the expression and subcellular localization of NO in muscle development and myoblast differentiation are largely unknown. In this study, we examined effects of the nitric oxide generated by a microwave plasma torch, on proliferation/differentiation of rat myoblastic L6 cells. Experimental data pertaining to nitric oxide production are presented in terms of the oxygen input in units of cubic centimetres per minute. The various levels of nitric oxide are observed depending on the flow rate of nitrogen gas, the ratio of oxygen gas, and the microwave power4. In order to evaluate the potential of nitric oxide as an activator of cell differentiation, we applied nitric oxide generated from the microwave plasma torch to L6 skeletal muscles. Differentiation of L6 cells into myotubes was significantly enhanced the differentiation after nitric oxide treatment. Nitric oxide treatment also increase the expression of myogenesis marker proteins and mRNA level, such as myogenin and myosin heavy chain (MHC), as well as cyclic guanosine monophosphate (cGMP), However during the myotube differentiation we found that NO activate oxidative stress signaling erks expression. Therefore, these results establish a role of NO and cGMP in regulating myoblast differentiation and elucidate their mechanism of action, providing a direct link with oxidative stress signalling, which is a key player in myogenesis. Based on these findings, nitric oxide generated by plasma can be used as a possible activator of cell differentiation and tissue regeneration.