• Title/Summary/Keyword: Protein A

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Protein Sequence Search based on N-gram Indexing

  • Hwang, Mi-Nyeong;Kim, Jin-Suk
    • Bioinformatics and Biosystems
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    • v.1 no.1
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    • pp.46-50
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    • 2006
  • According to the advancement of experimental techniques in molecular biology, genomic and protein sequence databases are increasing in size exponentially, and mean sequence lengths are also increasing. Because the sizes of these databases become larger, it is difficult to search similar sequences in biological databases with significant homologies to a query sequence. In this paper, we present the N-gram indexing method to retrieve similar sequences fast, precisely and comparably. This method regards a protein sequence as a text written in language of 20 amino acid codes, adapts N-gram tokens of fixed-length as its indexing scheme for sequence strings. After such tokens are indexed for all the sequences in the database, sequences can be searched with information retrieval algorithms. Using this new method, we have developed a protein sequence search system named as ProSeS (PROtein Sequence Search). ProSeS is a protein sequence analysis system which provides overall analysis results such as similar sequences with significant homologies, predicted subcellular locations of the query sequence, and major keywords extracted from annotations of similar sequences. We show experimentally that the N-gram indexing approach saves the retrieval time significantly, and that it is as accurate as current popular search tool BLAST.

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Partial characterization of a 17 kDa protein of Clonorchis sinensis

  • Chung, Young-Bae;Chung, Byung-Suk;Choi, Min-Ho;Chai, Jong-Yil;Hong, Sung-Tae
    • Parasites, Hosts and Diseases
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    • v.38 no.2
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    • pp.95-97
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    • 2000
  • A 17 kDa protein from Clonorchis sinensis adults was purified by a procedure including Sephacryl S-200 HR gel filtration and Q-Sepharose anion exchange chromatography. The protein was proved to be a cysteine protease as it showed hydrolytic activity toward Cbz-Phe-Arg-AMC in the presence of dithiothreitol and was inhibited by specific inhibitors such as iodoacetic acid or trans epoxy-succinly-L-leucyl-amido(4 guanidino) butane. The polyclonal antibody raised against the protein reacted to 17 kDa proteins of trematodes such as Paragonimuf westermani, Fasciola hepatica, Opisthorchis viverrini, Gymnophalloides seoi, and Metagonimus yokogawai. The antibody recognized the 17 kDa and 16 kDa cysteine proteases purified from C. sinensis, P. westemani, and G. seoi as well. These results suggest that the 17 kDa protein may be a cysteine protease commonly present in trematodes.

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A Study of the Anticoagulatory DNA from the Earthworm, Lumbricus rubellus, and its Regulatory DNA-Binding Protein

  • Kim, Gyoung-Mi;Yu, Kyoung-Hee;Woo, Jeong-Im;Bahk, Yun-Kyoung;Paik, Seung R.;Kim, Jung-Gyu;Chang, Chung-Soon
    • BMB Reports
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    • v.32 no.6
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    • pp.567-572
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    • 1999
  • We have previously shown that a DNA fragment is responsible for the anticoagulatory effect of an earthworm, Lumbricus rubellus. The anticoagluant increased the activated partial thromboplastin time (APTT) and also inhibited the thrombin activity observed with either N-${\alpha}$-p-tosyl-L-arginine methyl ester (TAME) or H-D-phenyl-alanyl-L-pipecoil-L-arginine-p-nitroanilide (S-2238). Since trypsin digestion of the anticoagulant further increased the APTT, the possible presence of a regulatory protein for the anticoagulatory DNA was investigated by digesting the anticoagulant with trypsin and isolating the DNA fragment with C4-reversed phase HPLC. The DNA fragment lacking a regulatory protein was eluted in the flow-through fraction, and analyzed with thrombin and activated factor X. Activated factor X activity was more strongly inhibited than thrombin activity. For DNA digestion, we treated the anticoagulant with DNase and purified the DNA-binding protein with a FPLC Resource-S cation exchange column. The regulatory protein, with an $M_r$ of 55.0 kDa, reduced the anticoagulatory effect of the DNA fragment.

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cDNA Cloning and Overexpression of an Isoperoxidase Gene from Korean-Radish, Raphanus sativus L.

  • Park, Jong-Hoon;Kim, Soung-Soo
    • BMB Reports
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    • v.29 no.2
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    • pp.137-141
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    • 1996
  • A partial cDNA encoding a Korean radish isoperoxidase was obtained from a cDNA library prepared from 9 day old radish root. In order to obtain Korean radish isoperoxidase cDNA, 5' RACE (rapid amplification cDNA end) PCR was performed and a cDNA (prxK1) encoding a complete structural protein was obtained by RT (reverse transcription)-PCR. Sequence analysis revealed that the length of the cDNA was 945 base pairs, and that of the mRNA transcript was ca. 1.6 kb. The deduced amino acid of the protein were composed of 315 amino acid residues and the protein was 92% homologous to turnip peroxidase, and 46% to 50% homologous to other known peroxidases. The 945 bp cDNA encoding Korean radish isoperoxidase was overexpressed in Escherichia coli up to approximately 9% of total cellular protein. The recombinant fusion protein exhibited 43 kDa on SDS-PAGE analysis and the activity level of the recombinant nonglycosylated protein was two fold higher in IPTG induced cell extracts than that of uninduced ones.

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Identification of a Protein Interacting with Human Nebulin SH3 Domain by Yeast Two-hybrid Screening

  • Lee, Min-A;Kim, Ji-Hee;Min, Byung-In;Park, Soo-Ho;Ko, Han-Suk;Kim, Chong-Rak
    • Biomedical Science Letters
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    • v.7 no.2
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    • pp.59-64
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    • 2001
  • Nebulin is an unusually large actin-binding protein specific to the skeletal muscle of vertebrates. The correlation of nebulin size with thin filament length have led to the suggestion that nebulin acts as a molecular ruler for the length of thin filaments. An SH3 domain occupies the C terminus of nebulin, in the sarcomeric Z-disk and is preceded by a 120-residue stretch containing multiple putative phosphorylation sites. SH3 domain mediates protein-protein interaction involved in the subcellular localization of proteins, cytoskeletal organization and signal transduction. However the binding partner and physiological role of nebulin SH3 domains remains unknown. Using the yeast two-hybrid system, we identified supervillin, an actin-binding protein, as a nebulin SH3 domain-interacting protein. The SH3 domain of nebulin binds to the sequence encoding amino acids 977 to 1335 of supervillin. But the sequence encoding amino acids 977 to 1335 displays weaker binding than the sequence encoding amino acids 977 to 1788.

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Purification and Identification of a Novel Antifungal Protein Secreted by Penicillium citrinum from the Southwest Indian Ocean

  • Wen, Chao;Guo, Wenbin;Chen, Xinhua
    • Journal of Microbiology and Biotechnology
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    • v.24 no.10
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    • pp.1337-1345
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    • 2014
  • A novel antifungal protein produced by the fungal strain Penicillium citrinum W1, which was isolated from a Southwest Indian Ocean sediment sample, was purified and characterized. The culture supernatant of P. citrinum W1 inhibited the mycelial growth of some plant pathogenic fungi. After saturation of P. citrinum W1 culture supernatants with ammonium sulfate and ion-exchange chromatography, an antifungal protein (PcPAF) was purified. The N-terminal amino acid sequence analysis showed that PcPAF might be an unknown antifungal protein. PcPAF displayed antifungal activity against Trichoderma viride, Fusarium oxysporum, Paecilomyces variotii, and Alternaria longipes at minimum inhibitory concentrations of 1.52, 6.08, 3.04, and $6.08{\mu}g/disc$, respectively. PcPAF possessed high thermostability and had a certain extent of protease and metal ion resistance. The results suggested that PcPAF may represent a novel antifungal protein with potential application in controlling plant pathogenic fungal infection.

BEAD BASED CHEMICAL REACTION SYSTEM USING TEMPERATURE AND FLUID CONTROL FOR CANCER DETECTION (유체와 온도 조절을 이용한 생화학 물질 반응용 마이크로칩의 개발)

  • Kim, Min-Su;Lee, Bo-Rahm;Yoon, Hyo-Jin;Kim, Byung-Gee;Lee, Yoon-Sik;Kim, Yong-Kweon
    • Proceedings of the KIEE Conference
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    • 2008.07a
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    • pp.1466-1467
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    • 2008
  • We describe here a novel micro total analysis system for the purification and identification of the affinity-captured proteins. Also we demonstrated the mass analysis of the Carcinoembrionic antigen (CEA) and Alpha femtoprotein which were chosen as the target cancer marker. For MALDI-TOF analyses, the proteins should to be separated from a protein mixture and be concentrated when needed. This procedure usually takes a long time even before protease-digested samples are to be obtained from them. Here, we describe integrated and efficient micro chip for protein purification and digestion for MALDI-TOF analyses. At first, disease protein is purified by passing the micro chamber from a protein mixture or human whole serum and released from the micro affinity beads by thermal heating. Purified protein is then transfer to the hole for trypsin digestion. The final sample is analyzed by MALDI-TOF. All the processes could be finished successfully within one hour, which renders MALDI-TOF analyses of a target protein quite simple.

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Concentration of Total Protein and Protein Fractions of Sera from Pregnant Rabbits (임신가토(姙娠家兎)의 혈청총단백량(血淸總蛋白量)과 단백분획상(蛋白分劃像)에 대(對)하여)

  • Jo, Chung Ho;Nam, Tchi Chou;Chang, Kyung Jin
    • Korean Journal of Veterinary Research
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    • v.14 no.1
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    • pp.29-31
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    • 1974
  • The total serum protein, protein fractions by paper electrophoresis and A/G ratio in pregnant rabbits were observed. The results obtained in this work were sumrnerized as follows: 1. Total serum protein and the fraction of serum albumin revealed a decrease with the advancing of gestation, especially total serum protein was decreased significantly on 3 weeks of pregnancy. There was a tendency to return tward control level on one week after delivery. 2. The fraction of ${\alpha}_1$, and ${\alpha}_2$-globulin showed little changes during the period of gestation. 3. The fraction of ${\beta}$-globulin was increased more or less during the period of gestation, and on one week after delivery showed considerable increase but the increase was statistically insignificant. 4. The fraction of ${\gamma}$-globulin revealed a variable changes during pregnancy but there was no significant differences. 5. A/G ratio was significantly decreased at 3 weeks of pregnancy and the ratio was near control level on one week after delivery.

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High-yield Expression and Characterization of Syndecan-4 Extracellular, Transmembrane and Cytoplasmic Domains

  • Choi, Sung-Sub;Kim, Ji-Sun;Song, Jooyoung;Kim, Yongae
    • Bulletin of the Korean Chemical Society
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    • v.34 no.4
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    • pp.1120-1126
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    • 2013
  • The syndecan family consists of four transmembrane heparan sulfate proteoglycans present in most cell types and each syndecan shares a common structure containing a heparan sulfate modified extracellular domain, a single transmembrane domain and a C-terminal cytoplasmic domain. To get a better understanding of the mechanism and function of syndecan-4 which is one of the syndecan family, it is crucial to investigate its three-dimensional structure. Unfortunately, it is difficult to prepare the peptide because it is membrane-bound protein that transverses the lipid bilayer of the cell membrane. Here, we optimize the expression, purification, and characterization of transmembrane, cytoplasmic and short extracellular domains of syndecan4 (syndecan-4 eTC). Syndecan-4 eTC was successfully obtained with high purity and yield from the M9 medium. The structural information of syndecan-4 eTC was investigated by MALDI-TOF mass (MS) spectrometry, circular dichroism (CD) spectroscopy, and nuclear magnetic resonance (NMR) spectroscopy. It was confirmed that syndecan-4 eTC had an ${\alpha}$-helical multimeric structure like transmembrane domain of syndecan-4 (syndecan-4 TM) in membrane environments.

Variation of Grain Traits in Landraces of Rice Collected from Vietnam (베트남 지방수집종 벼의 종실특성변이)

  • Kim, Hyun-Ho;Jo, Jae-Seong
    • Korean Journal of Agricultural Science
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    • v.24 no.2
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    • pp.121-125
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    • 1997
  • A total of 437 landraces of rice from Vietnam were analyzed for total seed protein by SDS-PAGE and phenol reaction. The different types of glutelin $\alpha$ subunits were detected. The level of wx protein with 60kDa molecular weight was divided into 3 groups, corresponding to non-glutinous, intermediate and glutinous starch types. Based on the variation in seed storage protein and wx protein, landraces were classified into 7 groups. Frequency distribution of types A and B of glutelin $\alpha$ subunits changed with the latitude at which rice landraces were collected. Geographical cline for phenol reaction was detected.

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