• 제목/요약/키워드: paramyosin

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전복 Paramyosin의 분리 및 그 성질 (PARAMYOSIN OF THE ABALONE, NOTOHALIOTIS DISCUS)

  • 변재형
    • 한국수산과학회지
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    • 제5권1호
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    • pp.29-38
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    • 1972
  • 복족류 근육단백질을 다른 연분동물의 그것과 비교생화학적으로 검토하고져 전복을 선정하여 근육단백질의 조성을 측정하고 그 주요 구성단백질인 paramyosin을 정열 단리하여 몇가지 생물물리화학적인 성질에 관하여 실험하였다. 전복근육의 단백조성은 수용성구분 $19\~22\%$, 염용성구분 $27\~39\%$, 알카리 가용성구분 $20\~26\%$, 그리고 stroma $20\~28\%$이었다. 염용성구분은 초원심분석에서 Paramyosin이 약 $65\%$, actomyosin이 약 $30\%$, myosin이 약$5\%$로서 이루어져 있음을 알았다. 그리고 초원심분석상 균질의 단일 표품으로 판명 분리된 본 실험에서의 전복 Paramyosin은 침항정수($S^{\circ}\;_{20,\;{\omega}$) 3.14s, 정전염용 $0.35{\mu}$ 이상, $25^{\circ}C$에서의 고유점도는 3.1이었고, 한편 동 Paramyosin은 염석분석에서 이 단백질의 염석절위가 $18\~30\%$이었다. 그리고 아미노산 분석결과, 구성아미노산은 arginine, aspartic acid, glutamic acid등이 많이 함유되어 있고, proline과 tryptophane이 흠여되어 있는 점 등 2매패류의 아미노산조성과 비슷하였으나, Iysine/arginine의 비는 0.61로서 2매패류보다 낮았다.

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방어 Myosin과 갈색띠 매물고둥 Paramyosin의 물리화학적 성질 (Physicochemical Characteristics of Yellowtail Myosin and Whelk Paramyosin)

  • 변재형;최영준
    • 한국식품과학회지
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    • 제19권1호
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    • pp.29-37
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    • 1987
  • 수산동물의 근육단백질 중 특히 많은 분포를 보이며. 생리적으로 뿐만 아니라 식품학적으로도 중요한 myosin과 paramyosin을 방어 보통육과 갈색띠 매물고둥에서 추출하여 각각의 물리 화학적 성질을 비교 검토 하였다. 방어 보통육 myosin은 분자량 $4.6{\times}10^5\;dalton$, 278 nm 에서의 분자흡광계수 $B^{1cm}_{1%}$ 5.44, 고유점도 1.60dl/g 및 그 소수성에 따른 형광강도는 740이었으며, 용해도에 비추어 등전점은 pH 5.0부근이었다. 한편 갈색띠 매물고둥 paramyosin은 분자량 $2{\times}10^5\;dalton$, 278nm 에서의 분자흡광계수$(B^{1cm}_{1%})$ 3.04, 고유점도 2.60dl/g 및 그 소수성에 의한 형광강도는 468이었으며, 등전점은 pH5.6부근이었다. 그리고 방어 myosin의 ${Ca}^{2+}-ATPase$ 활성은 0.342 ${\mu}moles-Pi/min/mg-protein$이었고, 분자당 40개의 SH기를 함유하고 있었다. 아미노산 조성의 분석 결과, 방어 보통육 myosin과 갈색띠 매물고둥 paramyosin의 총아미노산의 잔기수는 단백질 1 mole당 각각 4030잔기와 1694잔기이었으며, 극성 대 비극성 아미노산의 비율은 0.54와 0.47이었고, 산성 대 염기성 아미노산의 비율은 1.64와 2.42이었다.

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해산복족류(海産腹足類)의 Paramyosin의 분리(分離) 및 그 특성(特性)에 관(關)한 연구(硏究) (Isolation and Characterization of Paramyosins of Marine Gastropods)

  • 변재형
    • 한국식품영양과학회지
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    • 제2권1호
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    • pp.1-21
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    • 1973
  • The muscle of abalone, Notohaliotis discus (REEVE), and top-shell, Turbo cornutus Solander, were examined for protein composition. Then paramyosins which are known as one of the important structural protein of the muscle fibrils were isolated from the both muscle and their physico-chemical properties such as solubility, salting-out behaviour, intrinsic viscosity, ATPase activity, etc. involving amino acid composition and N-terminal amino acid residues were investigated to elucidate phylogenie characteristics more intensively from the viewpoint of comparative biochemistry. The analysis of protein composition resulted in the following estimations: abalone muscle; water-soluble protein of 22 %, salt-soluble protein, 34%, alkali-soluble protein, 20%, and stroma protein, 24%, and top-shell muscle; water-soluble protein of 16%, salt-soluble protein, 30%, alkali-soluble protein, 29%, and stroma protein, 25%, respectively. It is demonstrated in sedimentation analysis that paramyosin and myosin-actomyosin account for approximately 65% and 35% of the salt-soluble protein of abalone, and that the composition of both sediments in top-shell was approximately 70% and 30%, respectively. The ultracentrifugally homogenous paramyosins isolated essentially according to Bailey's ethanol-dried method from both of the muscle showed a $S^{\circ}_{20,w}$ of 3. 14s for abalone and a $S^{\circ}_{20,w}$ of 3.50s for top-shell. The both paramyosins were commonly rich in arginine, aspartic acid, and glutamic acid, while scarcely contained proline and tryptophan, in rough accord with the other paramyosins thus far reported. It is clear that these gastropod paramyosins showed of having the characteristic N-terminal amino acid residues such as N-aspartic acid, N-valine, N-serine, and N-threonine in common. The abalone paramyosin completely salted in with KCl beyond $0.35{\mu}$ and the top-shell paramyosin beyond $0.30{\mu}$. The abalone paramyosin was salted-out between 18% and 30% saturation of ammonium sulphate and the top-shell paramyosin between 22% and 29% saturation. The intrinsic viscosities at abalone and top-shell paramyosins at $25^{\circ}C$ were estimated respectively to be 3.1 dl/g and 2.6 dl/g showing somewhat higher than the values for some other paramyosins from lamellibranchs. In regard with the ATPase activity, the para myosin specimens did not exhibit any significant activity over through the pH conditions of 5 to 9.5. irrespective of the presence of $Ca^{++}$ or $Mg^{++}$. So was the case with the abalone paramyosin prepared by a slightly modified Bailey's wet-extraction method.

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Molecular Cloning and Characterization of a Paramyosin from Clonorchis sinensis

  • Park, Tae-Joon;Kang, Jung-Mi;Na, Byoung-Kuk;Sohn, Woon-Mok
    • Parasites, Hosts and Diseases
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    • 제47권4호
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    • pp.359-367
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    • 2009
  • Paramyosin is a myofibrillar protein present in helminth parasites and plays multifunctional roles in host-parasite interactions. In this study, we identified the gene encoding paramyosin of Clonorchis sinensis (CsPmy) and characterized biochemical and immunological properties of its recombinant protein. CsPmy showed a high level of sequence identity with paramyosin from other helminth parasites. Recombinant CsPmy (rCsPmy) expressed in bacteria had an approximate molecular weight of 100 kDa and bound both human collagen and complement 9. The protein was constitutively expressed in various developmental stages of the parasite. Imunofluorescence analysis revealed that CsPmy was mainly localized in the tegument, subtegumental muscles, and the muscle layer surrounding the intestine of the parasite. The rCsPmy showed high levels of positive reactions (74.6%, 56/75) against sera from patients with clonorchiasis. Immunization of experimental rats with rCsPmy evoked high levels of IgG production. These results collectively suggest that CsPmy is a multifunctional protein that not only contributes to the muscle layer structure but also to non-muscular functions in host-parasite interactions. Successful induction of host IgG production also suggests that CsPmy can be applied as a diagnostic antigen and/or vaccine candidate for clonorchiasis.

가열 변성에 따른 방어 Myosin과 갈색띠 매물고둥 Paramyosin의 소수성, 용해도, SH기 및 단백질간 상호작용의 변화 (Changes of Hydrophobicity, Solubility, SH Group and Protein-Protein Interaction in Yellowtail Myosin and Whelk Paramyosin During Thermal Denaturation)

  • 최영준;변재형
    • 한국식품과학회지
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    • 제19권2호
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    • pp.89-96
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    • 1987
  • 방어와 갈색띠 매물고둥에서 myosin과 paramyosin 을 추출하고, 이들 단백질의 가열중에 일어나는 변성기구를 아미노산 잔기와 SH기의 변화 및 단백질간의 상호작용 등을 측정하므로서 분석하였다. 각 단백질을 이루는 구성아미노산의 측쇄중 소수성 잔기의 유리정도는 가열온도 $65^{\circ}C$까지는 증가하였으나, 그 이상의 가열온도에서는 감소하는 경향을 보였다. 유리 소수성 잔기가 증가하여 감에 따라, 단백질간 상호작용도 활발하여 갔으며, 소수성 잔기의 유리정도가 감소하는 가열온도$65^{\circ}C$부터는 단백질의 응집이 일어나기 시작 하였다. 단백질간의 상호작용을 탁도로써 분석하여 Arrhenius식으로 해석한 결과, 방어 myosin은 3단계이상의 변성과정으로 구분할 수 있었으며, 갈색띠 매물고둥 paramyos은 2단계의 변성과정으로 구분할 수 있었다. 이들 두 단백질 소수성, 용해도, 유리 SH기의 수 및 단백질간의 상호작용 등은 온도함수와 밀접한 상관관계를 보였다.

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Mapping of the Complement C9 Binding Region on Clonorchis sinensis Paramyosin

  • Kang, Jung-Mi;Le, Huong Giang;Vo, Tuan Cuong;Yoo, Won Gi;Sohn, Woon-Mok;Na, Byoung-Kuk
    • Parasites, Hosts and Diseases
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    • 제60권4호
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    • pp.255-259
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    • 2022
  • Heliminthic paramyosin is a multifunctional protein that not only acts as a structural protein in muscle layers but as an immune-modulatory molecule interacting with the host immune system. Previously, we found that paramyosin from Clonorchis sinensis (CsPmy) is bound to human complement C9 protein (C9). To analyze the C9 binding region on CsPmy, overlapping recombinant fragments of CsPmy were produced and their binding activity to human C9 was investigated. The fragmental expression of CsPmy and C9 binding assays revealed that the C9 binding region was located at the C-terminus of CsPmy. Further analysis of the C-terminus of CsPmy to narrow the C9 binding region on CsPmy indicated that the region flanking 731Leu-780Leu was a potent C9 binding region. The CsPmy fragments corresponding to the region effectively inhibited human C9 polymerization. These results provide a precise molecular basis for CsPmy as a potent immunomodulator to evade host immune defenses by inhibiting complement attack.

Screening and Molecular Cloning of a Protective Antigen from the Midgut of Haemaphysalis longicornis

  • Hu, Yonghong;Zhang, Jincheng;Yang, Shujie;Wang, Hui;Zeng, Hua;Zhang, Tiantian;Liu, Jingze
    • Parasites, Hosts and Diseases
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    • 제51권3호
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    • pp.327-334
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    • 2013
  • Vaccination is considered a promising alternative for controlling tick infestations. Haemaphysalis longicornis midgut proteins separated by SDS-PAGE and transferred to polyvinylidene difluoride (PVDF) membrane were screened for protective value against bites. The western blot demonstrated the immunogenicity of 92 kDa protein (P92). The analysis of the P92 amino acid sequence by LC-MS/MS indicated that it was a H. longicornis paramyosin (Hl-Pmy). The full lenghth cDNA of Hl-Pmy was obtained by rapid amplification of cDNA ends (RACE) which consisted of 2,783 bp with a 161 bp 3' untranslated region. Sequence alignment of tick paramyosin (Pmy) showed that Hl-Pmy shared a high level of conservation among ticks. Comparison with the protective epitope sequence of other invertebrate Pmy, it was calculated that the protective epitope of Hl-Pmy was a peptide (LEEAEGSSETVVEMNKKRDTE) named LEE, which was close to the N-terminal of Hl-Pmy protein. The secondary structure analysis suggested that LEE had non-helical segments within an ${\alpha}$-helical structure. These results provide the basis for developing a vaccine against biting H. longicornis ticks.