• 제목/요약/키워드: N-terminal analysis

검색결과 485건 처리시간 0.036초

Nucleotide and Deduced Amino Acid Sequences of Rat Myosin Binding Protein H (MyBP-H)

  • Jung, Jae-Hoon;Oh, Ji-Hyun;Lee, Kyung-Lim
    • Archives of Pharmacal Research
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    • 제21권6호
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    • pp.712-717
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    • 1998
  • The complete nucleotide sequence of the cDNA clone encoding rat skeletal muscle myosin- binding protein H (MyBP-H) was determined and amino acid sequence was deduced from the nucleotide sequence (GenBank accession number AF077338). The full-length cDNA of 1782 base pairs(bp) contains a single open reading frame of 1454 bp encoding a rat MyBP-H protein of the predicted molecular mass 52.7kDa and includes the common consensus 1CA__TG' protein binding motif. The cDNA sequence of rat MyBP-H show 92%, 84% and 41% homology with those of mouse, human and chicken, respectively. The protein contains tandem internal motifs array (-FN III-Ig C2-FN III- Ig C2-) in the C-terminal region which resembles to the immunoglobulin superfamily C2 and fibronectin type III motifs. The amino acid sequence of the C-terminal Ig C2 was highly conserved among MyBPs family and other thick filament binding proteins, suggesting that the C-terminal Ig C2 might play an important role in its function. All proteins belonging to MyBP-H member contains `RKPS` sequence which is assumed to be cAMP- and cGMP-dependent protein kinase A phosphorylation site. Computer analysis of the primary sequence of rat MyBP-H predicted 11 protein kinase C (PKC)phosphorylation site, 7 casein kinase II (CK2) phosphorylation site and 4N-myristoylation site.

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A Solid State Controller for Self-Excited Induction Generator for Voltage Regulation, Harmonic Compensation and Load Balancing

  • Singh Bhim;Murthy S. S.;Gupta Sushma
    • Journal of Power Electronics
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    • 제5권2호
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    • pp.109-119
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    • 2005
  • This paper deals with the performance analysis of static compensator (STATCOM) based voltage regulator for self­excited induction generators (SEIGs) supplying balanced/unbalanced and linear/ non-linear loads. In practice, most of the loads are linear. But the presence of non-linear loads in some applications injects harmonics into the generating system. Because an SEIG is a weak isolated system, these harmonics have a great effect on its performance. Additionally, SEIG's offer poor voltage regulation and require an adjustable reactive power source to maintain a constant terminal voltage under a varying load. A three-phase insulated gate bipolar transistor (IGBT) based current controlled voltage source inverter (CC- VSI) known as STATCOM is used for harmonic elimination. It also provides the required reactive power an SEIG needs to maintain a constant terminal voltage under varying loads. A dynamic model of an SEIG-STATCOM system with the ability to simulate varying loads has been developed using a stationary d-q axes reference frame. This enables us to predict the behavior of the system under transient conditions. The simulated results show that by using a STATCOM based voltage regulator the SEIG terminal voltage can be maintained constant and free from harmonics under linear/non linear and balanced/unbalanced loads.

Pyruvate dehydrogenase phosphatase의 catalytical subunit의 구조와 활성에 대한 연구 (Structural and Functional Relationship of the Catalytical Subunit of Recombinant Pyruvate Dehydrogenase Phosphatase (rPDPc): Limited Proteolysis)

  • Kim, Young-Mi
    • Environmental Analysis Health and Toxicology
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    • 제17권1호
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    • pp.73-80
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    • 2002
  • Pyruvate dehydrogenase phosphatase (PDP)와 kinase는 당대사시 해당과정에서의 대사 산물인 pyruvate를 acetyl CoA로 만들어 구연산 회로로 진입시켜 주는 효소인 pyruvate dehydrogenase complex (PDC)의 활성을 조절하는 중요한 효소이다. PDP의 catalytic subunit는 PDC의 dihydrolipoamide acetyltransferase (E2), PDP regulatory subunit (PDPr), 그리고 칼슘 결합 도메인 등으로 구성되어 있는 것으로 추측되어지고 있다. 본 연구에서는 그 구조와 기능과의 상관관계를 알아보기 위해 PDPc를 E. coli JM101에서 발현시켜 순수 정제 후 단백분해 효소를 이용한 제한적 가수분해 방법을 이용해 그 구조와 기능과의 상관관계에 대해 연구하고자 하였다 정제된 PDPc는 trypsin, chymotrypsin, Arg-C 그리고 elastase를 이용하여 3$0^{\circ}C$ 그리고 pH 7.0에서 제한적으로 분해시켰으며 각 분해산물의 아미노 말단의 아미노산 배열을 분석하였다. 그 결과 PDPc는 trypsin, chymotrypsin, elastase에 의해 N-terminal의 50 kD과 C-terminal의 10 kD의 두개의 분해산물을 만들었으며, Arg-C에 의해 50kD의 분해산물은 약 35kD와 15kD으로 더 가수분해가 되었다. 이러한 결과로 볼 때 PDPc는 앞에서 추측한데로 세개의 주요한 기능적 도메인으로 이루어져 있음을 알 수 있었다 또한 C-terminal의 10kD은 PDPc의 활성에는 영향을 주지 않는 것으로 밝혀졌으나 다른 도메인의 기능은 더 연구가 되어져야 할 것으로 생각된다.

Euphorbia lathyris에서 분비되는 Latex 65kD 단백질의 특성규명 (Characterization of 65 kD Protein in Latex Excreted from Euphorbia lathyris)

  • 박희성
    • Journal of Plant Biotechnology
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    • 제31권4호
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    • pp.319-323
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    • 2004
  • Euphorbia lathyris의 유관세포로부터 분비되는 수용성 latex단백질을 10% SDS-polyacrylamide 전기영동으로 분리하여 특징적으로 잘 나타나고 있는 ELp65, ELp55, ELp43, ELp32 그리고 ELp23 등의 주요단백질을 확인하였다. 이들 latex 주요단백질들 중에서 ELp65는 ammonium sulfate 침전, gel permeation chromatography 그리고 ion exchange chromatography 등의 방법으로 순수 분리하였는데 ELp65의 N-terminal amino acid 서열분석에 의하면 이는 토마토의 p69a subtilisin-like pretense의 mature peptide의 앞부분과 강한 유사성을 지니고 있었으며 식물방어와 관련된 기능이 제시되었다. PCR증폭에 의하여 클로닝된 토마토의 p69a DNA를 probe로 이용하여 Southern blot hybridization을 수행한 결과 E. lathyris genome은 토마토의 subtilisin-like proteases와 유사한 정보를 지닐 수도 있는 3-5 유전자들로 구성된 gene family가 분석되었다.

INFLUNCE OF ELECTRO-MYOSCOPE(ACUSCOPE) ON INJURED QUADRICEPS FEMORIS WITH USING ELECTRO_MYOGRAPHIC ANALYSIS BUILT INTO THE ELCETROMYOSCOPE

  • 송병호
    • 대한물리치료과학회지
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    • 제7권2호
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    • pp.697-709
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    • 2000
  • 이 실험의 목적은 Electro-Myoscope 이 과다 사용에 의해 일어나는 대퇴사두 근육 통증에 미치는 효과에 대해 근전도를 이용한 연구입니다. 이를 위해 나이 15세부터 50세 사이의 18명의 여성 자원자들이 참여했다. 대퇴직근을 Cybex 운동기기를 사용해 Terminal knee extension 운동을 통해 과다사용으로 인한 통증이 유발되게 하였다. 그리고, Electro-Myoscope을 치료 후 근전도를 통해 대퇴사두근의 motor recruit 에 대해 기록 비교하였다. t-test 와 Chi square 분석 결과 대퇴사두근의 기능이 향상된 것으로 나타났다.

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Molecular Characterization of AceB, a Gene Encoding Malate Synthase in Corynebacterium glutamicum

  • Lee, Heung-Shick;Anthony J. Sinskey
    • Journal of Microbiology and Biotechnology
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    • 제4권4호
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    • pp.256-263
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    • 1994
  • The aceB gene, encoding for malate synthase, one of the key enzymes of glyoxylate bypass, was isolated from a pMT1-based Corynebacterium glutamicum gene library via complementation of an Escherichia coli aceB mutant on an acetate minimal medium. The aceB gene was closely linked to aceA, separated by 598 base pairs, and transcribed in divergent direction. The aceB expressed a protein product of Mr 83, 000 in Corynebacterium glutamicum which was unusually large compared with those of other malate synthases. A DNA-sequence analysis of the cloned DNA identified an open-reading frame of 2, 217 base pairs which encodes a protein with the molecular weight of 82, 311 comprising 739 aminoo acids. The putative protein product showed only limited amino acid-sequence homology to its counteliparts in other organisms. The N-terminal region of the protein, which shows no apparent homology with the known sequences of other malate synthases, appeared to be responsible for the protein s unusually large size. A potential calciumbinding domain of EF-hand structure found among eukaryotes was detected in the N-terminal region of the deduced protein.

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Expression, Purification and NMR studies of SH3YL1 SH3 domain

  • Shrestha, Pravesh;Yun, Ji-Hye;Lee, Weon-Tae
    • 한국자기공명학회논문지
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    • 제14권2호
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    • pp.105-116
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    • 2010
  • SH3YL1, a novel protein containing one Src homology 3 domain at the carboxyl terminus was first detected in mouse anagen skin cDNA. This protein had a significant homology with YHRO 16c/Ysc 84, the yeast Src homology 3 domain-containing protein. The sequence identity was remarkable at the carboxyl and amino-terminal Src homology 3 domain, suggesting that the novel protein is a mouse homolog of the yeast protein and thus was termed as SH3YL1. SH3YL1 is composed of two domains, a DUF500 at N-termini and a SH3 domain at C-termini. In our study we cloned the SH3 domain in bacterial expression system in Escherichia coli using pET32a vector with TEV protease cleavage site and purified as a monomer using affinity chromatography. The N-terminal poly-Histidine tag was cleaved with TEV protease and target protein was used for backbone studies. Our study showed that SH3 domain primarily consists of $\beta$-sheet which is in consistence with previous result performed on the truncated SH3 domain of SH3YL1.

Influence of Adaptor on the Calibration of Inductance Standards

  • Kassim, Dewi Mohd;Kim, Dan Bee;Kim, Wan-Seop
    • Journal of Electrical Engineering and Technology
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    • 제13권2호
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    • pp.911-917
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    • 2018
  • Influence of the adaptors on the calibration of $100{\mu}H$ inductance standard was studied as a function of torque, applied when tightening the standard inductor terminal with the adaptor. Two different homemade adaptors of BPO gold-plated brass (BPO-Au) and banana-copper (BN-Cu) were made for the connection between the LCR meter and the inductance standard. The measured inductance (L) of the standard inductor and the contact resistance ($R_C$) between the adaptor and the standard inductor terminal showed exponential decreases against the torque increase from $25cN{\cdot}m$ to $150cN{\cdot}m$. The measured L and the calculated equivalence series resistance ($R_S$) were dependent on the adaptor type as well as on the $R_C$. The results of the adaptor analysis imply that the BPO-Au adaptor with the lower $R_C$ is more suitable for the inductance calibration. The calculated inductance of $99.956{\mu}H$ corrected by subtraction of the adaptor inductance and the contact resistance contributions from the measured value using the BPO-Au adaptor agreed well with the certificate ($99.948{\mu}H$) of the PTB within the measurement uncertainty of $140{\mu}H/H$.

Molecular Cloning and Characterization of Calumenin in Rabbit Skeletal Sarcoplasmic Reticulum

  • Jung, Dai-Hyun;Kim, Do-Han
    • 한국생물물리학회:학술대회논문집
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    • 한국생물물리학회 2003년도 정기총회 및 학술발표회
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    • pp.53-53
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    • 2003
  • Calumenin was previously identified as a high affinity Ca$\^$2+/ binding protein in mouse cardiac sarcoplasmic reticulum (SR). For the present study, a 48 kDa skeletal homologue of calumenin was identified by sucrose-density gradient of rabbit skeletal SR membranes, concanavalin A treatment, 2D-gel electrophoresis, $\^$45/Ca$\^$2+/ overlay, Stains-all staining, and MALDI-TOF analysis. We attempted to clone the skeletal calumenin by RT-PCR based on mouse cardiac and human calumenin sequences. The deduced amino acid sequence (315 residues) of the skeletal calumenin showed high identity to mouse cardiac calumenin (90%). As seen in the cardiac calumenin, the deduced sequence contains a 19 amino acid N-terminal signal sequence and a HDEF C-terminal sequence, a putative retrieval signal to ER. Also, the skeletal calumenin contains one N-glycosylation site, three PKC phosphorylation sites, eight casein kinase 2 phosphorylation sites, and 6 EF-hand domains. GST-calumenin showed a conformational change and increased mobility in the presence of Ca$\^$2+/ in SDS-PAGE. Three calumenin interacting proteins (ryanodine receptor 1, glycogen phosphorylase, and phosphofructo kinase) were identified by pull-down assay with GST-calumenin and solubilized SR. All the interactions were Ca$\^$2+/dependent. The present results suggest that calumenin plays an important role in Ca$\^$2+/ homeostasis of muscle cells.

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Porphyromonas gingivalis, Prevotella intermedia, 그리고 Prevotella nigrescens에서의 hemin 결합 단백질에 대한 연구 (Isolation and Partial Characterization of Hemin-binding Cell Envelope Proteins from Porphyromonas gingivalis, Prevotella intermedia, and Prevotella nigrescens)

  • 김성조
    • Journal of Periodontal and Implant Science
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    • 제36권1호
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    • pp.155-165
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    • 2006
  • The results of this study confirm that the availability of hemin influences the expression of selected membrane proteins of Porphyromonas gingivalis, Prevotella intermedia, and Prevotella nigrescens. A 30 kDa (heated 24 kDa) hemin-binding protein whose expression is hemin regulated was identified and purified in P. gingivalis. A strong hemin-binding function was found by LDS-PAGE and TMBZ staining when P. gingivalis cells were grown under hemin-limited conditions. A 50 kDa cell envelope associated protein, whose expression is hemin regulated, is considered to be a putative hemin binding protein from P. intermedia and P. nigrescens, respectively. N-terminal amino acid sequence analysis of CNBr-digested 24 kDa hemin binding protein from P. gingivalis revealed that this protein belongs to a new, so far undescribed hemin-binding class of proteins. N-terminal amino acid sequence of a 50 kDa putative hemin binding protein from P. intermedia was identical with Enolase from Streptococcus intermedia. Work is in progress to further characterize the molecular structure of these proteins.