• 제목/요약/키워드: Myosin

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Review on the structural understanding of the 10S myosin II in the era of Cryo-electron microscopy

  • Anahita Vispi Bharda;Hyun Suk Jung
    • Applied Microscopy
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    • 제52권
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    • pp.9.1-9.5
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    • 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.

Myosin Head의 Spin Label이 온도 영향에 따른 구조 변화 연구 (Study of Structure Change by Temperature Effect in Spin Label of Myosin Head)

  • 김덕술;박근호
    • 한국응용과학기술학회지
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    • 제20권3호
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    • pp.268-273
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    • 2003
  • IASL(iodo acetamide spin label) and MSL(maleimide spin label) disordered the orderly helix arrangement of myosin in the rest state of spin level. Especially the effect of IASL was great. The muscle was isometrically tetanized with three trains of 3ms pulses every 50ms between $5^{\circ}C$ with $25^{\circ}C$. Equatorial reflection change inferred that myosin head was moved to the vicinity of actin filament by spin level. The intensity change of $143{\AA}$ and $72{\AA}$ could offer information of the mass projection of population of myosin head along the filament axis. The slope of intensity profile of the mass projection of $143{\AA}$ and reflection of IASL is appeared and that of MSL is appeared sharply. The decrease of $215{\AA}$ reflection intensity the periodical character of $143{\AA}$ reflection by spin label. The raise 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 by spin label effect. Because iodo acetamide has a tendency to decease the actin reflection, actin dose not bind myosin head. From this result, we can conclude that IASL and MSL are spin labeled on SH of myosin head and disordered the helix arrangement of actin.

Effects of Hydrostatic Pressure on Myofibrillar Protein Extracted from Bovine Semitendinosus

  • Lee, Eun-Jung;Kim, Yun-Ji;Lee, Nam-Hyouck;Yamamoto, Katsuhiro
    • 한국축산식품학회:학술대회논문집
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    • 한국축산식품학회 2004년도 정기총회 및 제33차 춘계 학술대회
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    • pp.198-201
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    • 2004
  • To investigate hydrostatic pressure (HP) effect on myofibrillar protein (Mf) extracted from bovine Semitendinosus muscle, Ca- and Mg-ATPase activities to evaluate denaturation of myosin and actin, and soluble protein contents were observed. In Mf treated with 100 MPa for 5 min was not observed denaturation of myosin and actin. In Mf treated with 200 MPa for 5 min, denaturation of myosin and actin were observed but inactivation rate was low (0.0136 $min^{-1}$). Inactivation rate of myosin and actin was dramatically increased above 300 MPa treatment. However denaturation of myosin and actin was not that critical with duration time. By increasing pressure size, the amount of myosin and actin in soluble protein eluted in 20 mM potassium phosphate buffer (pH 7.0) containing 0.6 M NaCl were decreased. SDS-PAGE of soluble protein released from Mf suspension in 0.1 M NaCl buffer (pH 7.0) showed that low molecular weight proteins (15${\sim}$36 KDa) were released by HP treatment above 200 MPa. From the results, denaturation of myosin and actin, and release of light molecule proteins of Mf were observed by HP treatment over 200 MPa.

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Molecular Identification and Expression of Myosin Light Chain in Shortspine Spurdog (Squalus mitsukurii)

  • Kim, Soo Cheol;Sumi, Kanij Rukshana;Sharker, Md Rajib;Kho, Kang Hee
    • 한국해양생명과학회지
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    • 제3권1호
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    • pp.1-8
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    • 2018
  • Myosin is considered as the vital motor protein in vertebrates and invertebrates. Our present study was conducted to decipher the occurrence of myosin in dog fish (Squalus mitsukurii). We isolated one clone containing 979 bp cDNA sequence, which consisted of a complete coding sequence of 453 bp and a deduced amino acid sequence of 150 amino acids from the open reading frame with molecular weight, isoelectric point and aliphatic index are 16.72 Kda, 4.49 and 78.00, respectively. It contained 428 bp long 3' UTR with single potential polyadenylation signals (AATAAA). The predicted EF CA2+ binding domains were identified in residue 6-41, 83-118 and 133-150. A BLAST search indicates this protein exhibits a strong similarity to whale shark (Rhincodon typus) MLC3 (91% identical) and also house mouse (Mus musculus) MLC isoform 3f (81% identical). Phylogenetic analysis revealed that this protein is a MLC 3 isoform like protein. This protein also demonstrates highly conserved region with other myosin proteins. Homology modeling of S. mitsukuri was performed using crystal structure of Gallus gallus skeletal muscle myosin II based on high similarity. Reverse transcription-polymerase chain reaction (PCR), quantitative PCR results exhibits dogfish myosin protein is highly expressed in muscle tissue.

닭고기의 근원섬유 단백질에 관한 연구 -2. 골격근 부위별로 추출한 근원섬유, 액토미오신 및 미오신의 ATPase 활성 비교- (Studies on the Myofibrillar Proteins from Chicken Muscle -2. Comparison of ATPase Activity in Myofibril, Actomyosin and Myosin Extracted from Leg and Pectoral Skeletal Muscle)

  • 박창식;공양숙;문윤희
    • 한국식품영양과학회지
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    • 제14권1호
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    • pp.82-87
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    • 1985
  • 닭의 가슴부위 및 다리부위의 골격근(骨格筋)에서 myofibril, actomyosin 및 myosin을 추출하고 ATPase activity(${\mu}mole$ pi/mg protein/min)로서 나타낸 몇가지 생물학적(生物學的) 활성(活性)을 비교하였다. 가슴부위에서 추출한 actomyosin, myofibril 그리고 myosin의 $Mg^{+2}$-ATPase 활성(活性)은 0.05M KCl에서 0.80, 0.42, 0.40으로서 다리부위에서 추출한 단백질(蛋白質)의 활성(活性)인 0.69, 0.33, 0.28 보다 높았다. 가슴부위와 다리부위의 myosin의 ATPase 활성(活性)은 EDTA 농도보다 $Mg^{+2}$농도가 높아지면서 ATPase 활성(活性)을 1/10정도 저해(沮害)시켰고, $Ca^{+2}$ 농도는 $10^{-3}M$에서 400%까지 활성(活性)을 증가시켰다. 가슴부위와 다리부위에서 추출한 actomyosin의 용해되는 시점(始點)은 각각 0.1M KCl 및 0.15 M KCl이었고 myosin인 경우는 각각 0.25 M KCi 및 0.30 M KCl이었다.

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Ardipusilloside-I stimulates gastrointestinal motility and phosphorylation of smooth muscle myosin by myosin light chain kinase

  • Xu, Zhili;Liang, Hanye;Zhang, Mingbo;Tao, Xiaojun;Dou, Deqiang;Hu, Liping;Kang, Tingguo
    • The Korean Journal of Physiology and Pharmacology
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    • 제21권6호
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    • pp.609-616
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    • 2017
  • Ardipusilloside-I is a natural triterpenoid saponin, which was isolated from Ardisia pusilla A. DC. The aim of the study was to evaluate the stimulation of ardipusilloside-I on gastrointestinal motility in vitro and in vivo. The experiment of smooth muscle contraction directly monitored the contractions of the isolated jejunal segment (IJS) in different contractile states, and the effects of ardipusilloside-I on myosin were measured in the presence of $Ca^{2+}$-calmodulin using the activities of 20 kDa myosin light chain ($MLC_{20}$) phosphorylation and myosin $Mg^{2+}$-ATPase. The effects of ardipusilloside-I on gastro emptying and intestinal transit in constipation-predominant rats were observed, and the MLCK expression in jejuna of constipated rats was determined by western blot. The results showed that, ardipusilloside-I increased the contractility of IJS in a dose-dependent manner and reversed the low contractile state (LCS) of IJS induced by low $Ca^{2+}$, adrenaline, and atropine respectively. There were synergistic effects on contractivity of IJS between ardipusilloside-I and ACh, high $Ca^{2+}$, and histamine, respectively. Ardipusilloside-I could stimulate the phosphorylation of $MLC_{20}$ and $Mg^{2+}$-ATPase activities of $Ca^{2+}$- dependent phosphorylated myosin. Ardipusilloside-I also stimulated the gastric emptying and intestinal transit in normal and constipated rats in vivo, respectively, and increased the MLCK expression in the jejuna of constipation-predominant rats. Briefly, the findings demonstrated that ardipusilloside-I could effectively excite gastrointestinal motility in vitro and in vivo.

Effects of the Myosin ATPase Inhibitor, 2,3-Butanedione-2-Monoxime, on Growth and Dimorphic Switches of Candida albicans

  • Woo, Mi-Young;Jwa, Mi-Ri;Kim, Jin-Mi;Song, Ki-Won
    • Journal of Microbiology and Biotechnology
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    • 제10권5호
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    • pp.606-611
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    • 2000
  • Dimorphic yeast Candida albicans reversibly switches between the form of yeast and hyphae depending on external conditions. We investigated possible roles of the myosin family in the growth and dimorphic switches of C. albicans with a general myosin ATPase inhibitor, 2,3-butanedione-2-monoxime (BDM). Transition to hyphae as well as proliferation by budding was completely inhibited by BDM at 16 mM. Presence of 16 mM BDM did not affect hyphae-to-bud transition but it blocked budding. The effects of BDM on yeast growth and dimorphic switches were reversible. More than 70% of the BDM-treated cells demonstrated defects in the amount and the polarized localization of F-actin as well as in the shape and migration of the nucleus, suggesting that myosin activities are needed in these cellular processes of C. albicans.

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연체류의 근원섬유단백질에 열 안정성에 관한 연구 (Studies on the Thermostability of Myofibrillar Proteins of Mollusca)

  • 신완철;송재철;김영호
    • 한국식품영양학회지
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    • 제10권2호
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    • pp.160-165
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    • 1997
  • 오징어의 actomyosin과 myosin은 3$0^{\circ}C$에서 최대 Vmax를 나타냈고 대합의 actomyosin과 myosin은 35$^{\circ}C$에서, HMM은 $25^{\circ}C$에서 최대 Vmax를 보였다. 또한 근원섬유단백질의 열 안정성은 염농도의 차이에 따라 크게 변하여 염농도가 높을수록 변성의 정도가 컸으며 대합의 근원섬유단백질이 오징어의 근원섬유단백질보다 높은 열 안정성을 나타냈다. 근원섬유단백질에 3%의 에탄올을 첨가하여 가온하면 변성은 가속화되었으며 변성속도에 있어서도 동물간에 차이가 있는 것으로 나타났다.

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Identification of the Interaction between Rat Translationally Controlled Tumor Protein/IgE-dependent Histamine Releasing Factor and Myosin Light Chain

  • Kim, Min-Jeong;Jung, Jae-Hoon;Choi, Eung-Chil;Park, Hae-Young;Lee, Kyung-Lim
    • BMB Reports
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    • 제34권6호
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    • pp.526-530
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    • 2001
  • The translationally controlled tumor protein (TCTP), also known as the IgE-dependent histamine releasing factor (HRF), was used in the yeast two-hybrid system to screen the interacting molecules. We obtained the N-terminus truncated rat fast myosin alkai light chain from the rat skeletal muscle cDNA library in the screening. Since either TCTP/HRF or the myosin light chain is known to be associated with histamine secretion from RBL-2H3 cells, we investigated the possible interaction between rat TCTP/HRF and nonmuscle myosin light chain in these cells. We used affinity chromatography and coimmunoprecipitation. Our data suggests that HRF and the myosin light chain interact, which may play an important role in histamine release in RBL-2H3 cells.

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