• Title/Summary/Keyword: Elastase-like serine protease

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Molecular Cloning, Expression Analysis and Enzymatic Characterization of Elastase-like Serine Protease from the Olive Flounder (Paralichthys olivaceus) (넙치로부터의 Serine Protease의 분자생물학적 클로닝, 발현, 특성분석)

  • Han, Jin-Wook;Seo, Jung-Soo;Ahn, Sang-Jung;Lee, Jin-Young;Park, Ju-Hyeon;Kim, Na-Young;Kim, Moo-Sang;Hwang, Jee-Youn;Chung, Joon-Ki;Lee, Hyung-Ho
    • Journal of Fisheries and Marine Sciences Education
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    • v.26 no.4
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    • pp.808-822
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    • 2014
  • 넙치 (Paralichthys olivaceus)로부터 elastase-like serine protease (PoElSp)를 암호화하는 cDNA를 클로닝하여 그 서열을 분석한 결과, PoElSp 유전자는 269 아미노산을 암호화하는 978염기쌍으로 구성되었다. PoElSp 유전자의 조직 특이적 발현 양상을 RT-PCR법으로 조사한 결과, 간, 비장 및 소장에서 그 발현이 크게 나타났다. lipopolysaccharide (LPS)로 인위적 세균감염을 유도한 후, 1시간째에 콩팥에서, 3시간째에는 근육에서, PoElSp 유전자의 발현이 크게 증가하였다. 또한, 이 유전자의 발현은 비장에서 LPS 주입 후 1-24시간동안 점차로 증가하였다. pro-mature PoElSp (proPoElSp)에 해당하는 cDNA를 pET32a 벡터 시스템을 이용하여 대장균에서 발현시켰다. 이 재조합 proPoElSp 단백질의 활성은 gelatin zymography 방법과 합성형광 Z-Phe-Arg-AMC의 분해법을 이용하여 측정하였다. 단백질 분해효소 활성을 위한 최적 pH는 7.5였다. 실험결과들을 종합하면, PoElSp 단백질은 면역 반응에서 중추적 역할을 하리라 판단된다.

Aptamers (nucleic acid ligands) for trypsin-like serine proteases

  • Gal, Sang-Wan;Jeong, Yong-Kee;Satoshi Nishikawa
    • Journal of Life Science
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    • v.12 no.1
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    • pp.14-18
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    • 2002
  • Subpopulations of nucleotides that bind specifically to a variety of proteins have been isolated from a population of random sequence RNA/DNA molecules. Roughly one in $10^{13}$ random sequence RNA/DNA molecules folds in such a way as to create a specific binding site for small ligands. Since the development of in vitro selection procedure, more than 50 nucleic acid ligands (aptamers) have been isolated. These molecules are very useful for the study of molecular recognition between nucleic acid and protein/organic compound. In addition to these basic studies this method gives us a dream to produce new drugs against several diseases. We focused on several aptamers which specifically binds to trypsin-like serine proteases (thrombin, human neutrophil elastase, activated protein C and NS3 protease of human hepatitis C virus) and want to introduce their structural characteristics and some functions.

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Proteomic analysis of porcine pancreas development

  • Choi, Jong-Soon;Cho, Young-Keun;Yoon, Sung-Ho;Kwon, Sang-Oh;Koo, Deog-Bon;Yu, Kweon
    • BMB Reports
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    • v.42 no.10
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    • pp.661-666
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    • 2009
  • Porcine pancreas development is not well studied at the molecular level despite being a therapeutic resource for diabetic patients. In this study, we investigated expression of lineage markers and performed proteomic analysis. Expression of the early lineage markers Pdx1 and Ptf1a was developmentally conserved between mice and pigs, whereas expression of the islet differentiation marker Pax4 was delayed in porcine compared with murine pancreas development. Proteomic analysis found that expression levels of chymotrypsinogen were down-regulated during porcine pancreas development while those of digestive enzymes like lipases, elastase and serine protease were up-regulated. In addition, specific isoforms of protein folding assistants such as protein disulfide isomerase and prefoldin were expressed at specific stages during the maturation of digestive enzymes. Taken together, these results show that development of the porcine pancreas is regulated by a concerted interplay of pancreas lineage marker proteins and other specified proteins, resulting in a functional endocrine and exocrine organ.