• Applied Microscopy
• /
• 제52권
• /
• pp.9.1-9.5
• /
• 2022

The compact smooth muscle 10S myosin II is a type of a monomer with folded tail and the heads bending back to interact with each other. This inactivated form is associated with regulatory and enzymatic activities affecting myosin processivity with actin filaments as well as ATPase activity. Phosphorylation by RLC can however, shuttle myosin from the inhibited 10S state to an activated 6S state, dictating the equilibrium. Multiple studies contributed by TEM have provided insights in the structural understanding of the 10S form. However, it is only recently that the true potential of Cryo-EM in deciphering the intramolecular interactions of 10S myosin state has been realized. This has led to an influx of new revelations on the 10S inactivation, unfolding mechanism and association in various diseases. This study reviews the gradual development in the structural interpretation of 10S species from TEM to Cryo-EM era. Furthermore, we discuss the utility of Cryo-EM in future myosin 10S studies and its contribution to human health.

• 생명과학회지
• /
• 제9권6호
• /
• pp.646-652
• /
• 1999

The effect of temperature on the structure change of the SH of myosin head have been investigated with improved resolution by x-ray diffraction using synchrotron radiation. The movement of myosin head and conformational change of contractile molecules were occurred in the muscle contraction. IASL (iodo acetamide) and MSL (maleimide) disordered the orderly helix arrangement of myosin in the rest state of spin level. The temperature effect on the structure change was great at the UL in the equatorial reflection. But those of IASL and MSL were minor. Equatorial reflection (10, 11) change inferred that myosin head was moved to the vicinity of actin filament by temperature change (from $25^{\circ}C$ to $0^{\circ}C$) at UL, but spin level was not changed. The intensity change of 143 $\AA$ and 72 $\AA$ could offer information of the mass profection of population of myosin heads along the filament axis. The slope of intensity profile of the mass profection of 143$\AA$ and reflection of MSL is appeared sharply and those of UL and IASL were not changed. The decrease of MSL actin reflection at 51 $\AA$ and 59 $\AA$ in the actin reflection change refers that the shifted myosin head binds a certain actin or changes an actin structure. From these results, we could conclude that IASL and MSL were spin labeled on SH of myosin head and disordered the helix arrangement of actin.

• 한국임상수의학회지
• /
• 제32권1호
• /
• pp.62-68
• /
• 2015

There is no reliable indicator available for the diagnosis of horse laminitis, although the disease is common and costly. This study was performed to develop a specific diagnostic biomarker for laminitis. We have identified 33 differentially expressed proteins in plasma of a horse suffering laminitis that is experimentally induced by an overdose of oligofructose, in comparison with normal horse plasma. Among the proteins, myosin-9 mRNA was found in RNA sequencing analysis to be expressed specifically in laminitis tissues compared to other horse tissues. It is thus suggested that expression of plasma myosin-9 may be used for the diagnosis of equine laminitis.

• Asian-Australasian Journal of Animal Sciences
• /
• 제16권9호
• /
• pp.1384-1389
• /
• 2003

In order to clarify one of the biological functions of pork, we investigated whether a peptic hydrolysate of denatured porcine crude myosin showed inhibitory activity against angiotensin I-converting enzyme (ACE), which contributed to hypertension. Our results indicated that this hydrolysate showed relatively strong activity, and we therefore attempted to separate the involved peptides, which were considered to be active substances. To isolate these active peptides, the hydrolysate was separated using a solidphase separation, gel filtration high-performance liquid chromatography (HPLC), and two kinds of reverse phase HPLC. In each stage of separation, many fractions were detected, almost all of which showed ACE inhibitory activity. Thus, we suggested that the activity of the hydrolysate as a whole was a result of the activities of the many individual peptides. Six peaks were distinguished, with yields from 34 to 596 ppm of original crude myosin. In addition to the six peaks, many other active fractions were found throughout the separation steps, strongly suggesting that whole porcine crude myosin itself had ACE inhibitory activity. Moreover, pork as food was considered to function as an ACE inhibitory material in vivo, because pork proteins consist primarily of crude myosin, which included almost all the myofibrillar structural proteins.

• 한국식품과학회지
• /
• 제30권4호
• /
• pp.862-870
• /
• 1998

Myosin과 그 subfragment인 S-1과 LMM를 항원으로 하는 간접경합 효소면역분석법(Ci-ELISA)을 이용하여 동결육의 변화를 연구함으로써 수입 냉동육과 국내산 냉장 한우육과의 차별화를 위한 기초 자료를 마련하기 위하여 본 연구를 실시하였다. 각 항체의 표준곡선을 작성하였으며 최적반응범위는 $4{\sim}125{\;}{\mu}g/mL$이었고, 검출한계는 $0.1{\;}{\mu}g/mL$이었다. 확립된 Ci-ELISA를 이용하여 각각 $-10^{\circ}C,{\;}-20^{\circ}C,{\;}-50^{\circ}C$, 그리고 $-80^{\circ}C$에서 동결 기간 동안의 변화와 해동과 재동결에 따른 변화를 측정하였을 때 동결 기간 중 anti-MWM IgG와 반응한 myosin은 비교구와 비교하였을 때 $-20^{\circ}C$에서 변성도가 가장 컸으며 $-50^{\circ}C$에서 가장 적게 변성되었다. 단백질 용해성의 변화와 myosin과 항체와의 면역 친화성을 비교하였을 때 용해성이 감소하는 속도보다 더 빠르게 myosin이 변성되는 것으로 판단되었다. Anti S-1 IgG의 면역 친화성의 변화는 anti-MWM IgG에서 얻은 결과와 비교하였을 때 전혀 다른 경향을 보였다. anti-MWM IgG는 면역 친화성이 1개월 이후 급격히 감소된 데 비하여 anti S-1 IgG는 그와 같은 큰 변화를 나타내지 않았다. 반복되는 해동과 재동결 처리에서 anti-MWM IgG는 myosin과의 반응에서 2회 해동시부터 공격한 반응성의 감소를 보였으며 6회 해동시에는 85%까지 반응력을 상실했다(P<0.05). 두 실험 조건에서 $-20^{\circ}C$의 처리구의 myosin이 가장 심한 영향을 받은 것으로 나타났다. Anti-LMM IgG는 anti-MWM IgG에서 얻은 결과와 매우 유사한 경향을 보였다. $-10^{\circ}C$와 $-20^{\circ}C$에서 처리된 우육의 myosin은 저온에서 처리된 시료의 그것보다 보다 약 $10{\sim}15%$ 정도 더 많은 변성을 보인 것으로 사료된다.

• International Journal of Industrial Entomology and Biomaterials
• /
• 제9권1호
• /
• pp.127-130
• /
• 2004

We describe here the cloning and characterization of a cDNA encoding a putative myosin light chain-2 (MLC-2) from the mole cricket, Gryllotalpa orientalis. The G. orientalis MLC-2 cDNA sequences comprised of 615 bp with 205 amino acid residues with a calculated molecular weight of approximately 23 kDa. The deduced protein sequence of G. orientalis MLC-2 cDNA showed 64% and 54% identity to Drosophila melanogaster MLC-2 and D. yakuba MLC-2, respectively. Northern blot analysis confirmed the muscle-specific expression of G. orientalis MLC-2.

• 한국응용과학기술학회지
• /
• 제24권4호
• /
• pp.362-368
• /
• 2007

To study the relationship between elementary biochemical states and structural states of the actomyosin crossbridges in muscle, the effects of binding of MgADP to myosin heads in the rigor muscle were examined by X-ray diffraction using synchrotron radiation. X-ray diffraction studies have been made to investigate the effects of binding of ADP on the structure of glycerinated rabbit skeletal muscle in the rigor state. The intensity increase was accompanied by a slight but distinct decrease in the 5.9 am layer-line intensity close to the meridian. These results strongly suggest that myosin heads altered their attached conformation in the proximal end toward the plane perpendicular to the fiber axis when MgADP was bound to them. We found that the intensity of the 14.5 nm-based meridional reflections increase by 20-50% when MgADP was added to the rigor muscle in the presence of hexokinase and myokinase inhibitor.

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

• 한국수산과학회지
• /
• 제5권2호
• /
• pp.57-62
• /
• 1972

잉어의 배육 골격근에서 actomyosin을 추출하여 몇 가지 생물물리화학적인 성질을 측정 검토한 결과는 다음과 같다. 즉, actamyosin의 침강 정수($s_{20},\;_\omega$)는 24.76s이고 그 점성은 단백 농도의 증가와 더불어 급상승하여 다른 actomyosin의 그것과 비숫한 경향을 보였다. 그리고, ATP-sensitivity는 $147\~176$이었고, PH영향하에서 ATPase activity는 산성측이 6,0부근, 알칼리성측이 9.5부근이었으므로 ATPase activating site가 산, 알칼리 양측에 있음을 알았다.

• 생명과학회지
• /
• 제21권8호
• /
• pp.1076-1082
• /
• 2011

미세소관(microtubule) 위를 이동하는 키네신은 분비소포를 이동시키는 운동단백질이다. KIF5s (KIF5A, KIF5B and KIF5C)는 세포막으로 싸인 각종 세포 내 소기관과 결합하여 미세소관을 따라 목적지까지 이동시킨다는 결과는 알려져 있지만, 어떻게 상대의 cargo를 인식하는지는 밝혀지지 않았다. 본 연구는 KIF5B의 결합 단백질을 동정하기 위하여 효모 two-hybrid system을 사용하여 KIF5B와 특이적으로 결합하는 Myo9b을 확인하였다. Myo9b는 액틴위를 이동하는 운동단백질로 다른 KIF5s들과도 결합함을 효모 two-hybrid assay로 확인하였다. 또한 Myo9s의 GTPase 활성화 단백질(GAP) 영역은 KIF5B와 결합하는데 필수영역임을 확인하였고, 이러한 단백질간의 결합은 Glutathione S-transferase (GST) pull-down assay를 통하여서도 확인하였다. 생쥐의 뇌 파쇄액에 KIF5B들의 항체로 면역침강을 행하여 Myo9s 단백질을 확인한 결과, KIF5s는 Myo9s 단백질과 특이적으로 함께 침강하였다. 이러한 결과들은 kinesin-I는 액틴 결합 운동단백질과 직접 결합함을 보여준다.