• Applied Biological Chemistry
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• 제39권2호
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• pp.104-111
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• 1996

감자의 genomic DNA library로 부터 분리한 proteinase inhibitor II (pin2) 유전자들의 제한효소 지도를 작성 하였던바 이미 분리된 상처 유발 (wound-inducible)pin2K 유전자의 것과 상이성이 있는 pin2T를 분리하여 염기서열을 결정하였다. 두 유전자의 염기서열은 전체적으로 약 86%의 동일성을 보였으며 특히 promoter 부위의 염기서열은 pin2K 유전자의 전사개시 부위의 상대적인 위치인 -714까지 네부분의 결손(20 내지 60bp)을 제외하던 약 91%수준의동일성을 보였다. 분리한 유전자들의 promoter 부위를 표지 유전자인 CAT와 GUS 유전자에 연결 시킨후 담배에서의 발현을 추적하였던바, pin2K 유전자의 promoter에 의한 표지유전자의 발현은 상처에 의해 발현 되었으나 pin2T 유전자의 promoter에 의한 표지유전자의 발현은 상처 유무와 관계없이 낮은 수준으로 나타났다. 또한 pin2T 유전자의 Promoter 내의 결손은 핵 단백질의 promoter에의 결합에 영향을 주지 않았으며 상처 유발pin2K 유전자의 promoter 염기서열과 비교하였을때 pin2T 유전자의 promoter 부위내에 5'-AGTAAA-3'라는 특별한 염기부위가 자연적으로 제거된것을 알수 있었다. 또한 5'-AGTAAh-3'의 염기부위가 다른 상처 유발 유전자들에서는 흔히 발견되고, 다른 식물 유전자들의 Promoter에서는 쉽게 발견이 되지 않았다. 따라서 상처 유발 pin2K 유전자의 Promoter내에 상처 유발과 관련있는 특별한 염기부위가 자연적으로 결실되어 pin2T 유전자의 발현이 상처 유발성을 잃은것으로 짐작된다.

• Parasites, Hosts and Diseases
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• 제30권3호
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• pp.191-200
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• 1992

한국산 유혈목이에서 스파르가눔 충체를 수집하고, 이들 충체의 추출액에서 ion-exchange chromatography와 affinity chromatography를 실시하여 cysteine proteinase를 순수 정제하였다. 경제된 효소의 최적 pH는 5.5이었고, 최적 mole 농도는 0.IM (0.1M sodium acetate, pH5.5) 이었다. 정제된 대소는 thiol-dependent이고, $4^{\circ}C$에서 pH 5.0일 때 24시간 동안 안전성을 보였다. 효소의 환성도는 저분자 합성기질인 CBZ-phe-arg-AFC에 대 해 활성이 높았다. 정제된 효소는 척추동물의 산성 cysteine proteinase의 억제인자에 감수성을 보였다. UItrogel AcA54 column chromatography로 정제된 cysteine proteinase의 분자량을 측정한 결과 28,000 dalton이었다. 정제된 효소는 collagen type I과 hemoglobin을 분해하였다. Immunoblot한 결과 정제된 효소는 스파르가눔증 환자의 혈청과 반응하였다. 이상의 결과에서 스파르가눔의 cysteine proteinase는숙주 체내이동, 조직침수성 및 영양소 섭취에 관여할 것이라 추정되며, 정제된 효소는 스파르가눔 현중의 혈청학적 진단에 이용될 수 있을 것으로 생각된다.

• Parasites, Hosts and Diseases
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• 제44권4호
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• pp.321-330
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• 2006

The pathogenic mechanism of granulomatous amebic encephalitis (GAE) and amebic keratitis (AK) by Acanthamoeba has yet to be clarified. Pretense has been recognized to play an important role in the pathogenesis of GAE and AK. In the present study, we have compared specific activity and cytopathic effects (CPE) of purified 33 kDa serine proteinases from Acanthamoeba strains with different degree of virulence (A. healyi OC-3A, A. lugdunensis KA/E2, and A. castelianii Neff). Trophozoites of the 3 strains revealed different degrees of CPE on human corneal epithelial (HCE) cells. The effect was remarkably reduced by adding phenylmethylsulfonylfluoride (PMSF), a serine proteinase inhibitor. This result indicated that PMSF-susceptible proteinase is the main component causing cytopathy to HCE cells by Acanthamoeba. The purified 33 kDa serine proteinase showed strong activity toward HCE cells and extracellular matrix proteins. The purified proteinase from OC-3A, the most virulent strain, demonstrated the highest enzyme activity compared to KA/E2, an ocular isolate, and Neff, a soil isolate. Polyclonal antibodies against the purified 33 kDa serine proteinase inhibit almost completely the proteolytic activity of culture supernatant of Acanthamoeba. In line with these results, the 33 kDa serine proteinase is suggested to play an important role in pathogenesis and to be the main component of virulence factor of Acanthamoeba.

• 대한흉부심장혈관외과학회:학술대회논문집
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• 대한흉부외과학회 1993년도 제25차년차학술대회 및 총회
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• pp.46-46
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• 1993

• Journal of Chest Surgery
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• 제28권3호
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• pp.221-227
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• 1995

Malignancy is one of the several exogenous and endogenous factors that increase serum alpha 1-PI. In fact, serum levels of alpha 1-PI were significantly elevated in the patients with the nonresectable bronchogenic cancer. the purpose of this work was to determine if the immediate postoperative change of serum alpha 1-PI level following tumor resection relates to the patient`s postoperative course. Clinical experimental study was carried out to investigate the postoperative changes of serum alpha 1-PI level following operation for 20 cases of bronchogenic cancer and 10 cases of control, nephrectomy patients Alpha 1-PI concentrations in serum was quantitated by use of radial immunodiffusion technique.The results were as follows ; Preoperative serum level of alpha 1-PI was significantly elevated in patients with bronchogenic cancers [p < 0.001 , when compared to normal control levels. Immediate postoperative serum alpha 1-PI level was significantly increased in patients with bronchogenic cancer [p < 0.05 , but slightly decreased at control groups. The peak serum level of alpha 1-PI was the postoperative three days, and then gradually decreased at the 5, 9, 14 days, but slightly elevated comparing to preoperative alpha 1-PI levels. Serum alpha 1-PI level in patients with adenocarcinoma was elevated, when compared to squamous cell carcinoma, but not significantly. According to the stages of the bronchogenic cancer, each levels of the serum alpha 1-PI were slightly different, but the whole postoperative changes were the general similarity. There were no significant difference in changes of the serum alpha 1-PI level, according to the operative procedures. As the alpha 1-PI is acute reactant, that it was required at the reoperative state of the bronchogenic cancer and rapid response, consumption or requirement were occurred, postoperatively. Therefore, alpha 1-PI can be perioperative indicator for the evaluation of the bronchogenic cancer.

• Parasites, Hosts and Diseases
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• 제40권2호
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• pp.93-99
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• 2002

The effect of a secretory proteinase from the pathogenic amoebae Acanthamoeba castellanii on host's defense-oriented or regulatory proteins such as immunoglobulins, interleukin-1, and protease inhibitors was investigated. The enzyme was found to degrade secretory immunoglobulin A (slgA), IgG, and IgM. It also degraded $interleukin-1{\alpha}$ ($IL-l{\alpha}$) and $IL-l{\beta}$. Its activity was not inhibited by endogenous protease inhibitors, such as ${\alpha}$2-macroglobulin, ${\alpha}l-trypsin$ inhibitor, and ${\alpha}2-antiplasmin$. Furthermore, the enzyme rapidly degraded those endogenous protease inhibitors as well. The degradation of host's defense-oriented or regulatory proteins by the Acanthanoeba proteinase suggested that the enzyme might be an important virulence factor in the pathogenesis of Acanthamoeba infection.

• 한국균학회지
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• 제14권2호
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• pp.101-107
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• 1986

The properties of carboxyl proteinase which was contained in Poria cocos (Schw.) Wolf were investigated by means of the purification with 0.65 ammonium sulfate saturation, DEAE cellulose and Sephadex G-75 gel filtration. This enzyme was found to hydrolyze only peptide bond between glutamyl-L-tyrosine of carbobenzoxy-L-glutamyl-L-tyrosine among the synthetic substrates of carbobenzoxy-L-glutamyl-L-tyrosine, hippuryl- L-phenylalanine and hippuryl-L-arginine. This enzyme was inhibited by $Zn^{+2},\;Fe^{+2},\;Ca^{+2},\;CN^{-1},\;P_2O_7^{-4}$ ions, but stimulated by $Hg^{+2}$ ion. Also, this enzyme was inhibited by organic compounds such as L-lysine, L-phenylalanine, hippuryl-L-phenylalanine, diazoacetyl-DL-norleucine methyl ester (DAN) and 1,2-epoxy-3-(P-nitrophenoxy)propane(EPNP). In particular, the activity was inhibited by L-lysine till 20 minutes of preincubation time rapidly, and by DAN in the presence of $Cu^{+2}$ ion more rapidly after 30 minutes than DAN in the absence of $Cu^{+2}$ ion. L-Lysine was found to be a competitive inhibitor and its $K_i$ value was determined to be 0.12 mmole by Dixon plot.

• 한국식물생명공학회:학술대회논문집
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• 한국식물생명공학회 2003년도 추계학술대회 발표논문 초록집
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• pp.97-97
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• 2003

• Journal of Plant Biotechnology
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• 제6권4호
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• pp.145-145
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• 2004

• 한국식물생명공학회:학술대회논문집
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• 한국식물생명공학회 2002년도 춘계학술대회
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• pp.169-169
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• 2002