• 제목/요약/키워드: Amino acids and Proteins Analysis

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생물정보학을 이용한 연체동물의 NLS (Nuclear Localization Signals) 포함 단백질의 분석 (Bioinformatic Analysis of NLS (Nuclear Localization Signals)-containing Proteins from Mollusks)

  • 이용석;강세원;조용훈;곽희철;채성화;최상행;안인영;박홍석;한연수;고원규
    • 한국패류학회지
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    • 제22권2호
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    • pp.109-113
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    • 2006
  • 연체동물 유래 아미노산 서열 22,138 개에서 NLS가 예측되는 아미노산 서열은 266 개였으며 이는 연체동물 전체 아미노산 중 1.2% 정도였다. 또한 현재 등재되어 있는 연체동물 8,314 종 중 NLS를 포함한 아미노산이 밝혀진 생물은 60여종에 불과 하였다. 현재 알려진 연체동물 서열 중에는 두족 강의 경우가 NLS를 포함한 아미노산이 많을 것으로 예측되었다.

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방어 보통육과 혈합육의 단백질 및 아미노산조성의 사후변화 (Changes Occurred in Protein and Amino Acid Compositions during Postmortem Aging of White and Dark Muscle of Yellowtail at $2^{\circ}C$)

  • 김장양;최영준;변재형
    • 한국수산과학회지
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    • 제15권2호
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    • pp.123-136
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    • 1982
  • 방어의 배육 보통육과 측육 혈합육을 $2^{\circ}C$에 보장하여 두고 선도변화 총계별로 단백질의 조성과 각축의 구성 및 유리아미노산의 조성을 분석하였으며, 건향질단백질과 근원섬유단백질에 대하여는 구성단백질의 분포변화를 확인하기 위하여 $NaDodSO_4$변한 다음, polyacrylamide겔 전기영동상을 비교 해석함으로써 적색육어류의 선도변화와 보통육 및 혈합육의 단백질조성 변화와의 관련성을 검토하였다. K-값과 휘발성염기질소 및 pH값으로 판정했을 때, 혈합육은 보통육에 비하여 선도변화가 빨랐다. 사후 선도변화와 더불어 근원질단백질과 노원섬유단백질은 감소하는 반면, 세포내잔사단백질은 상대적으로 증가하였으며, 이러한 변화는 보통육에 비하여 혈합육이 훨씬 심한 편이었다. Sodium dodecyl sulfate 화한 단백질을 polyacrylamide겔 전기영동함으로 분석한 결과, 보통육의 근원질단백질은 16개 성분, 그리고 혈합육의 근원질단백질은 12개 성분으로 구성되어 있었으며, 보장 10일째부터 보통육의 근원질단백질은에는 41,000dalton과 18,000dalton의 양분이 조금씩 증가하였고, 26,000 dalton과 23,500 dalton 및 23,000 dalton되 각 성분은 점차 감소하였다. 혈합육의 근원질단백질에 있어서는 49,000 dalton의 성분은 감소하였고 47.000 dalton의 성분은 새로이 형성되었으며, 26,000 dalton의 성분은 소감하였다. 근원섬유단백질의 전기영동분석결과에 의하면, 보통육의 근원섬유단백질은 17개 성분, 그리고 혈합육의 근원조직단백질은 16개의 성분으로 구성되어 있었다. 보장 10일에부터 보통육의 근원섬유단백질중에는 40,000 dalton의 성분이 증가하였고, 37,500 dalton의 성분은 점차 감소하였으며, 32,000 dalton의 성분은 새로히 출현하였다. 혈합육의 근원섬유단백질은 보장 9일째부터 58,000 dalton과 64,000 dalton의 성분은 그 농도가 단가하였으며, 17,500의 light chain-2 단백질은 소감하였고, 32,000 dalton의 성분은 새로이 출현하였다. 이 같은 전기영동상의 변화는 근원질단백질이 근원섬유단백질에 비하여 빨랐으며, 근원섬유단백질의 경화에 있어서는 혈합육쪽이 보통육에 비하여 빨랐다. 양 육의 단백질을 구성하는 아미노산을 분석한 결과, 즉살한 방어의 보통육 단백질은 글루탐산, 아스팔트산, 류신. 알기닌, 알라닌 등은 그 양이 어느 정도 많았고, 프롤린과 트?토판은 그 양이 적었으며, 혈합육에 있어서도 비슷한 경형이었다. 그리고 10일이 경과한 보통육에 있어서는 글루탐산과 아스팔트산이 현저히 감소하였으며, 9일이 경과한 혈합육에 있어서는 알라닌, 글리신, 그리고 알기닌이 현저히 감소하였다. 유리아미노산의 조성을 분석 결과, 즉살한 방어의 보통육중에는 히스티딘이 총유리아미노산의 약 $63\%$를, 그리고 혈합육중에는 타우린이 총유리아미노산의 약 $67\%$를 차지하는 양을 보였다. 그리고 $2^{\circ}C$에서 10일이 경과했을 때, 보통육에서는 히스티딘, 발린, 타우린이 증가하였으며, 알라닌, 류신 및 글리신 등은 조금 감소하였다. 같은 온도에서 9일이 경과한 혈합육에서는 타우린, 페닐알라닌, 글리신은 증하였으며, 히스티딘, 알라닌, 세린 등은 감소하였다.

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Brucella abortus 국내 분리주의 Heat Shock Protein 암호 groE 유전자의 염기서열 분석과 발현 (Sequence analysis and expression of groE gene encoding heat shock proteins of Brucella abortus isolates)

  • 김태용;김지영;장경수;김명철;박창식;한홍율;전무형
    • 대한수의학회지
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    • 제45권1호
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    • pp.45-53
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    • 2005
  • GroE that is a heat shock protein composed of GroEL and GroES is known as an immunodominant target of both the humoral and cellular immune responses in bovine brucellosis. This study was carried out to characterize groE gene encoding heat shock proteins of B. abortus isolated in Korea and to evaluate the immunogenicity of the GroE protein expressed in E. coli system. In PCR the specific signals with the size of 2,077 bp were detected in five strains isolated from the mammary lymphnodes of the dairy cattle that were serologically positive and the reference strains. In comparison of the sequences of nucleotides and amino acids among the strains, GroES showed 100% identity in both sequences. GroEL was evaluated 99.0~99.9% in nucleotides and 98.0~100% homology in amino acids. The groE gene including groES and groEL was inserted into pET29a vector and constructed pET29a-GroE recombinant plasmids. The inserted groE was confirmed by digestion with Nco1 and EcoR1 endonucleases and nucleotide sequencing. E. coli BL (DE3) was transformed with pET29a-GroE, named as E. coli BL (DE3)/pET29a-GroE. In SDS-PAGE, it was evident that the recombinant plasmid effectively expressed the polypeptides for GroES (10 kDa) and GroEL (60 kDa) in 0.5, 1 and 2 hours after IPTG induction. The immuno-reactivity of the expressed proteins were proved in mouse inoculation and Western blot analysis.

대두 펩타이드의 아미노산 조성 및 평균소수도가 흰쥐의 혈청 콜레스레롤 농도에 미치는 영향 (Effects of Amino Acid Composition and Average Hydrophobicity of Soybean Peptides on the Concentration of Serum Cholesterol in Rats)

  • 한응수;이형주;손동화
    • 한국식품과학회지
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    • 제25권5호
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    • pp.552-557
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    • 1993
  • 대두 펩타이드의 아미노산 조성 및 평균소수도가 혈청 콜레스테롤의 농도에 미치는 영향을 조사하기 위하여, 대두 단백질(ISP), 카세인(CNP), 이들 단백질을 펩신으로 가수분해하여 pH에 다른 펩타이드 침전 획분들(SHT, SH8, SH6, SH4, CHT) 및 각 단백질과 같은 조성의 아미노산 혼합물(SAA, CAA)을 흰쥐에 섭취시키고 혈청콜레스테롤 농도를 측정하였다. 각 펩타이드의 아미노산 조성을 분석하고, 이로부터 구한 Lys/Arg 비율 및 소수성 아미노산의 몰분율은 혈청 콜레스테롤의 농도와 상관관계가 없었으며, Tanford, Manavalan 및 Meirovitch법으로 구한 평균소수도와도 상관관계가 없었다. 그러나, Krigbaum법으로 구한 평균소수도와는 상관계수가 -0.736으로 음의 상관관계이어서, 식이 펩타이드의 krigbaum 평균소수도가 높을 수록 래트의 혈청 콜레스테롤 농도가 낮아졌다.

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Identification and Characterization of a Conserved Baculoviral Structural Protein ODVP-6E/ODV-E56 from Choristoneura fumiferana Granulovirus

  • Rashidan, Kianoush Khajeh;Nassoury, Nasha;Giannopoulos, Paresa N.;Guertin, Claude
    • BMB Reports
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    • 제35권6호
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    • pp.595-603
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    • 2002
  • A gene that encodes a homologue to baculoviral ODVP-6E/ODV-E56, a baculoviral envelope-associated viral structural protein, has been identified and sequenced on the genome of Choristoneura fumiferana granulovirus (ChfuGV). The ChfuGV odvp-6e/odv-e56 gene was located on an 11-kb BamHI subgenomic fragment using different sets of degenerated primers, which were designed using the results of the protein sequencing of a major 39 kDa structural protein that is associated with the occlusion-derived virus (ODV). The gene has a 1062 nucleotide (nt) open-reading frame (ORF) that encodes a protein with 353 amino acids with a predicated molecular mass of 38.5 kDa. The amino acid sequence data that was derived from the nucleotide sequence in ChfuGV was compared to those of other baculoviruses. ChfuGV ODVP-6E/ODV-E56, along with othe baculoviral ODVP-6E/ODV-E56 proteins, all contained two putative transmembrane domains at their C-terminus. Several putative N-and O-glycosylation, N-myristoylation, and phosphorylation sites were detected in the ChfuGV ODVP-6E/ODV-E56 protein. A similar pattern was detected when a hydrophobicity-plots comparison was performed on ChfuGV ODVP-6E/ODV-E56 with other baculoviral homologue proteins. At the nucleotide level, a late promoter motif (GTAAG) was located at -14 nt upstream to the start codon of the GhfuGV odvp-6e/odv-e56 gene. a slight variant of the polyadenylation signal, AATAAT, was detected at the position +10 nt that is downstream from the termination signal. A phylogenetic tree for baculoviral ODVP-6E/ODV-E56 was constructed using a maximum parsimony analysis. The phylogenetic estimation demonstrated that ChfuGV ODVP-6E/ODV-E56 is most closely related to those of Cydia pomonella granulovirus (CpGV) and Plutella xylostella granulovirus (PxGV).

인삼으로부터 Acyl-CoA-binding Protein 유전자의 동정 및 계통적 분석 (Isolation and Phylogenetic Analysis of Acyl-CoA-binding Protein Gene from Panax ginseng C.A. Meyer)

  • 인준교;류명현;최광태;최관삼;김세영;양덕춘
    • 식물조직배양학회지
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    • 제28권4호
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    • pp.201-204
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    • 2001
  • Acyl-binding protein (ACBP)은 긴사슬 acyl-CoA와 결합하는 고도로 보존되어 있는 세포질 단백질이다. 인삼의 유용 유전자를 대량으로 분석하기 위하여 제작된 인삼 모상근cDNA library로부터 인삼 ACBP유전자가 분리되었다. 이 유전자는 길이가 453 bp이고 264 bp의 open reading frame (10kDa)을 가지고 있었다. 인삼 ACBP의 아미노산 서열을 다른 식물체에서 보고된 것과 비교한 결과 castor bean과 89.5%로 매우 높은 유사성을 나타내었으며, lilly, Digitalis. Arabidopsis, rape 등과 각각 81.8%, 80.7%, 73%, 71.9%의 유사성을 나타내었다. 그러나 인삼의 ACBP는 Arabidopsis 와 rape의 ACBP보다 5개의 아미노산이 적은 87개의 아미노산으로 이루어져 있었고 어떠한 signal peptide로 발견되지 않았다. 그리고 현재 보고되어 있는 다른 식물체의 ACBP와 계통분석을 한 결과 인삼의 ACBP는 Arabidopsis나 cotton 보다는 castor bean과 매우 가까운 유연관계에 있는 것으로 나타났다.

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Analysis of Symptom Determinant of Cucumber mosaic virus RNA3 via Pseudorecombinant Virus in Zucchini Squash

  • Choi, Seung-Kook;Yoon, Ju-Yeon;Choi, Jang-Kyung;Kim, Kook-Hyung;Sohn, Seong-Han
    • The Plant Pathology Journal
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    • 제23권4호
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    • pp.272-280
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    • 2007
  • Isolates of Cucumber mosaic virus (CMV) collected in Korea, were compared with their pathological features in tobacco and zucchini squash. Full-length cDNA clone of RNA3 was generated by using long-distance RT-PCR. Transcript RNA3 from the cDNA clone was inoculated onto host plants with transcripts RNA1 and RNA2 of Fny strain, generating RNA3-pseudorecombinant CMV. Timing and severity of systemic symptom was not significantly different among the pseudorecombinant CMVs in tobacco, compared with strains Fny-CMV and Pf-CMV. However, the pseudorecombinant CMVs induced two different systemic symptoms (mosaic vs. chlorotic spot) in zucchini squash. Based on symptom induction, the pseudorecombinant CMVs were categorized into two classes. The severity and timing of symptoms were correlated with viral RNA accumulations in systemic leaves of zucchini squash, suggesting that different kinetics of virus movement associated with CMV proteins are crucial for systemic infection and symptom development in zucchini squash. The analysis of movement proteins (MP) of CMV strains showed high sequence homology, but the differences of several amino acids were found in the C-terminal region between Class-I-CMV and Class-II-CMV. The analysis of coat proteins (CP) showed that the CMV isolates tested belonged to CMV subgroup I and the viruses shared overall 87-99% sequence identity in their genomes. Phylogenetic analysis of MP and CP suggested that biological properties of Korean CMV isolates have relationships associated with host species.

The Dynamics of Protein Decomposition in Lakes of Different Trophic Status - Reflections on the Assessment of the Real Proteolytic Activity In Situ

  • Siuda, Waldemar;Kiersztyn, Bartosz;Chrost, Ryszard J.
    • Journal of Microbiology and Biotechnology
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    • 제17권6호
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    • pp.897-904
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    • 2007
  • The aim of this paper is to discuss the methodology of our investigation of the dynamics of protein degradation and the total in situ protealytic activity in meso/eutrophic, eutrophic, and hypereutrophic freshwater environments. Analysis of the kinetics and rates of enzymatic release of amino acids in water samples preserved with sodium azide allows determination of the concentrations of labile proteins $(C_{LAB})$, and their half-life time $(T_{1/2})$. Moreover, it gives more realistic information on resultant activity in situ $(V_{T1/2})$ of ecto- and extracellular proteases that are responsible for the biological degradation of these compounds. Although the results provided by the proposed method are general y well correlated with those obtained by classical procedures, they better characterize the dynamics of protein degradation processes, especially in eutrophic or hypereutrophic lakes. In these environments, processes of protein decomposition occur mainly on the particles and depend primarily on a metabolic activity of seston-attached bacteria. The method was tested in three lakes. The different degree of eutrophication of these lakes was clearly demonstrated by the measured real proteolytic pattern and confirmed by conventional trophic state determinants.

ZAS3 represses NFκB-dependent transcription by direct competition for DNA binding

  • Hong, Joung-Woo;Wu, Lai-Chu
    • BMB Reports
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    • 제43권12호
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    • pp.807-812
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    • 2010
  • $NF{\kappa}B$ and ZAS3 are transcription factors that control important cellular processes including immunity, cell survival and apoptosis. Although both proteins bind the ${\kappa}B$-motif, they produce opposite physiological consequences; $NF{\kappa}B$ activates transcription, promotes cell growth and is often found to be constitutively expressed in cancer cells, while ZAS3 generally represses transcription, inhibits cell proliferation and is downregulated in some cancers. Here, we show that ZAS3 inhibits $NF{\kappa}B$-dependent transcription by competing with $NF{\kappa}B$ for the ${\kappa}B$-motif. Transient transfection studies show that N-terminal 645 amino acids is sufficient to repress transcription activated by $NF{\kappa}B$, and that the identical region also possesses intrinsic repression activity to inhibit basal transcription from a promoter. Finally, in vitro DNA-protein interaction analysis shows that ZAS3 is able to displace $NF{\kappa}B$ by competing with $NF{\kappa}B$ for the ${\kappa}B$-motif. It is conceivable that ZAS3 has therapeutic potential for controlling aberrant activation of $NF{\kappa}B$ in various diseases.

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.