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

• BMB Reports
• /
• 제48권1호
• /
• pp.48-53
• /
• 2015

Oxidized LDL (oxLDL) performs critical roles in atherosclerosis by inducing macrophage foam cell formation and promoting inflammation. There have been reports showing that oxLDL modulates macrophage cytoskeletal functions for oxLDL uptake and trapping, however, the precise mechanism has not been clearly elucidated. Our study examined the effect of oxLDL on non-muscle myosin heavy chain IIA (MHC-IIA) in macrophages. We demonstrated that oxLDL induces phosphorylation of MHC-IIA (Ser1917) in peritoneal macrophages from wild-type mice and THP-1, a human monocytic cell line, but not in macrophages deficient for CD36, a scavenger receptor for oxLDL. Protein kinase C (PKC) inhibitor-treated macrophages did not undergo the oxLDL-induced MHC-IIA phosphorylation. Our immunoprecipitation revealed that oxLDL increased physical association between PKC and MHC-IIA, supporting the role of PKC in this process. We conclude that oxLDL via CD36 induces PKC-mediated MHC-IIA (Ser1917) phosphorylation and this may affect oxLDL-induced functions of macrophages involved in atherosclerosis.

• 한방재활의학과학회지
• /
• 제19권1호
• /
• pp.11-21
• /
• 2009

Objectives : This study was performed to investigate the effects of Semen Persicae(SP) on muscle type change and vascularization effect by measuring expression of Myosin heavy chain (MHC) and Vascular endothelial growth factor (VEGF) protein in the Middle cerebral artery occlusion(MCAo) rats. Methods : The middle cerebral artery was occluded, SP extraction was administrated for 4 days. The effects of SP on muscle type change and vascularization were measured. Results : 1. VEGF protein expressions in the Semen Persicae oral administration group of MCAo group were increased compared to the control group. 2. There were no significant difference between the Semen Persicae oral administration of MCAo group and the control group in MHC isoform (Type I, Type IIa) expression change. Conclusions : The present study demonstrates the effect of Semen Persicae in the vascularizing after ischemia, but has no significant effect in musle type change and the improvement of musle atrophy.

• Animal cells and systems
• /
• 제14권1호
• /
• pp.25-30
• /
• 2010

We employed a new and improved differential display reverse transcription-polymerase chain reaction (DDRT-PCR) method, which involves annealing control primers (ACPs), to identify the genes that are specifically or prominently expressed in olive flounder (Paralichthys olivaceus) juveniles (35 days post-hatch; dph) compared to larval-stage (dph 21) flounder. Using 60 ACPs, we identified eight differentially expressed genes (DEGs) and basic local alignment search tool (BLAST) searches revealed eight known genes. Gene expression levels were confirmed by RT-PCR. Phosphoglucose isomerase (PGI) was highly expressed at 21 dph, while nephrosin, myosin light chain (MLC), myosin heavy chain (MHC), carboxypeptidase A, chymotrypsin B, fish-egg protein, and matrix protein were expressed at 35 dph. PGI, MLC, and MHC expression was further analyzed by RT-PCR. The differentially expressed genes identified in this study may provide insights into the molecular basis of development in olive flounder.

• 한국축산식품학회지
• /
• 제36권6호
• /
• pp.819-828
• /
• 2016

Muscle fiber characteristics account for meat quality and muscle fibers are mainly classified into three or more types according to their contractile and metabolic properties. However, the majority of previous studies on bovine skeletal muscle are based on myosin ATPase activity. In the present study, the differences in the characteristics of muscle fibers classified by the expression of myosin heavy chain (MHC) among four bovine skeletal muscles such as longissimus thoracis (LT), psoas major (PM), semimembranosus (SM) and semi-tendinosus (ST) and their relationships to beef quality were investigated. MHCs 2x, 2a and slow were identified by LC-MS/MS and IIX, IIA and I fiber types were classified. PM, which had the smallest size and highest density of fibers regardless of type, showed the highest myoglobin content, CIE $L^*$, $a^*$, $b^*$ and sarcomere length (p<0.05), whereas ST with the highest composition of IIX, showed high shear force and low sarcomere length (p<0.05). The correlation coefficients between muscle fiber characteristics and meat quality showed that type IIX is closely related to poor beef quality and that a high density of small-sized fibers is related to redness and tenderness. Therefore, the differences in meat quality between muscles can be explained by the differences in muscle fiber characteristics, and especially, the muscles with good quality are composed of more small-sized fibers regardless of fiber type.

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

• 한국식품과학회지
• /
• 제22권6호
• /
• pp.640-645
• /
• 1990

우육에 7.5%의 당을 첨가하고 $121^{\circ}C$에서 가열처리하여 근원섬유단백질의 전기영동 pattern 변화를 알아보았다. 당을 첨가하지 않은 경우 가열처리시간이 길어짐에 따라 myosin heavy chain(MHC) 등의 분자량이 큰 단백질의 band강도는 크게 약화되었지만 actin 등의 분자량이 작은 단백질은 거의 변화가 없었으며 당을 첨가한 처리구에서는 가열처리에 의하여 MHC 등의 band강도가 더욱 약화되었는데 특히 glucose 첨가시 15분간 가열처리구에서는 actin band의 강도도 크게 약해졌다. 시료에 peptidase를 작용시킨 후 추출한 단백질의 전기영동 pattern은 거의 변화가 없었으며 chymotrypsin을 작용시켰을 때 근원섬유를 구성하는 단백질들이 소화된 것을 보여주었으나 갈변반응이 심하게 발생하였던 glucose 첨가구에서는 myosin분자 등의 중합체가 나타났다 Trypsin을 작용시켰을 때 전체적으로 chymotrypsin의 경우보다 더욱 소화된 것을 보여주면서 glucose 첨가구에서는 myosin분자 등의 중합체가 더욱 강하게 나타났으며 peptidase chymotrypsin, trypsin을 동시에 작용시켜도 glucose 첨가구에서는 myosin 분자 등의 중합체가 소화되지 못하며 그 강도도 fructose 첨가구의 것보다 강한 것을 보여 주었다.

• 한국식품과학회지
• /
• 제28권2호
• /
• pp.292-297
• /
• 1996

본 연구에서는 오징어를 주원료로_하여 제조한 식해의 숙성과정증 수분함량 및 속성온도에 따른 단백질화학적 변화를 검토하였다. 속성과정중 pH의 변화는 수분함량 및 숙성온토가 높을 수록 빠르게 저하되었으며 이에 따른 산도의 변화도 빠르게 증가되었다. 또한 젖산균의 생성속도에 있어서도 수분함량 및 숙성온도가 높을수록 빨랐다. 한편 숙성과정중의 오징어 근단백질의 subunit 조성의 변화는 식해의 제조과정중 MHC에 상당하는 성분은 완전히 소실되었으며, 그 후 숙성기간이 경과함에 따라 A에 상당하는 성분도 서서히 소실되었다. 이와같은 결과는 적어도 숙성초기가지는 오징어육의 자가분해효소에 의한 것으로 숙성초기까지는 오징어의 자가분해효소에 의한 것으로 사료되며, 그후의 단백질의 분해과정은 숙성 진행과 함께 숙성 관여미생물 성장에 의해 생성되는 protease 및 오징어육 자체내 산성 protease에 의한 것으로 생각되었다.

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

• Asian-Australasian Journal of Animal Sciences
• /
• 제19권1호
• /
• pp.102-108
• /
• 2006

Microbial transglutaminase (MTGase) was investigated to determine whether it was an effective binding agent for the processing of low-quality pork loins. MTGase especially promoted the coagulation of myosin heavy chain (MHC). However, the effect of MTGase on MHC from low-quality meat was less than that from the normal meat when the reaction time was not enough. The breaking strength of the heat-induced gel made of myosin B from low-quality meat with MTGase was lower than that of normal meat. Sausage made with low-quality meat with MTGase did not exhibit improved hardness, as compared to that made with normal meat. Results of this study indicated that use of low-quality meat in the manufacture of sausage was feasible to get textural property equal to that of normal meat sausage, when a half or more of the raw material was normal meat and MTGase was used in the sausage.