• 한국미생물·생명공학회지
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• 제20권3호
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• pp.257-262
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• 1992

Streptomyces virginiae가 생산하는 virginimycin 생산 유도인자(virginiae butanolide C,VB-C) 결합 단백질의 ligand(VB-C)와의 결합활성에 미치는 일반적인 성질을 검토한 결과, 본 VB-C 결합단백질은 막성분을 제외한 세포질에 90% 이상 존재하며, 최적 pH는 7.0인 것으로 입증되었다. KCL 존재하 약 15%의 결합활성이 증대되었으며,$Mo^{6+}$ 이온 존재시 60%의 결합활성 저하를 보였다.

• BMB Reports
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• 제38권3호
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• pp.266-274
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• 2005

High temperature requirement A (HtrA) and its homologues constitute the HtrA familiy proteins, a group of heat shock-induced serine proteases. Bacterial HtrA proteins perform crucial functions with regard to protein quality control in the periplasmic space, functioning as both molecular chaperones and proteases. In contrast to other bacterial quality control proteins, including ClpXP, ClpAP, and HslUV, HtrA proteins contain no regulatory components or ATP binding domains. Thus, they are commonly referred to as ATP-independent chaperone proteases. Whereas the function of ATP-dependent chaperone-proteases is regulated by ATP hydrolysis, HtrA exhibits a PDZ domain and a temperature-dependent switch mechanism, which effects the change in its function from molecular chaperone to protease. This mechanism is also related to substrate recognition and the fine control of its function. Structural and biochemical analyses of the three HtrA proteins, DegP, DegQ, and DegS, have provided us with clues as to the functional regulation of HtrA proteins, as well as their roles in protein quality control at atomic scales. The objective of this brief review is to discuss some of the recent studies which have been conducted regarding the structure and function of these HtrA proteins, and to compare their roles in the context of protein quality control.

• 한국동물학회지
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• 제39권2호
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• pp.215-222
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• 1996

In the previous studies, we have demonstrated that prorenin converting enzyme (PRECE) was identical to the epidermal grouch factor-binding protein (EGF-BP) type B, which was a member of the mouse glandular kallikrein family, To examine whether or not EGF-BP type A and C are involved in the processing of prorenin, we have cloned the CDNAS of the EGF-BP type h and C from a library of male ICR mouse submandibular gland (SMGI. And then CHO cells were transfected with the EGF-BP expression plasmids. and stable cell lines expressing a high level of the EGF-BPS precursor were obtained. The conditioned medium was then treated with trypsin, which has been knotvn to effectively convert the EGF-BP type A and C precursor to the active forms. 수ubsequentlv, the prorenin converting activity of the trypsin-treated or untreated medium was examined. PRECE converted exactly prorenin to renin, but the prorenin converting activities of EGF-BP type A and C were not detected. From these results, it seems that only type B of these EGF-BPs is involved in processing Ren 2 prorenin in mouse SMG.

• Journal of Microbiology
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• 제37권1호
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• pp.14-20
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• 1999

Two extracellular proteases, EP I and EP II, from cells of Oligotropha carboxydovorans (formerly Pseudomonas carboxydovorans) DSM 1227 grown in nutrient broth were purified to greater than 95% homogeneity in five steps using azocasein as a substrate. The final specific activities of EPs I and II were 214.9 and 667.4 units per mg of protein. The molecular weights of native EPs I and II were determined to be 23,000. Sodium dodecyl sulfate-gel electrophoresis revealed the two enzymes to be monomers. The enzymes were found to be serine-type proteases. The activity of EP I was stimulated by Ca2+, Mg2+, and Ba2+, but that of EP II was not. The enzymes were completely inhibited by Fe2+, Hg2+, Co2+, Zn2+, and Cd2+. EDTA and EGTA exhibited a strong inhibitory effect on EP I. The optimal pH for the two enzymes was pH 9.0. The optimal temperatures for EP I and II were 60 and 50$^{\circ}C$, respectively. The enzymes were stable under alkaline conditions. The thermal stability of EP I was higher than that of EP II. Cell-free extracts did not inhibit the purified enzymes. The enzymes were active on casein, azocasein, azocoll, and carbon monoxide dehydrogenase, but weakly active with bovine serum albumin.

• Clinical and Experimental Reproductive Medicine
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• 제48권4호
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• pp.273-282
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• 2021

Since December 2019, severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) has spread rapidly, resulting in a pandemic. The virus enters host cells through angiotensin-converting enzyme 2 (ACE2) and transmembrane protease serine subtype 2 (TMPRSS2). These enzymes are widely expressed in reproductive organs; hence, coronavirus disease 2019 (COVID-19) could also impact human reproduction. Current evidence suggests that sperm cells may provide an inadequate environment for the virus to penetrate and spread. Oocytes within antral follicles are surrounded by cumulus cells, which rarely express ACE2 and TMPRSS2. Thus, the possibility of transmission of the virus through sexual intercourse and assisted reproductive techniques seems unlikely. Early human embryos express coronavirus entry receptors and proteases, implying that human embryos are potentially vulnerable to SARS-CoV-2 in the early stages of development. Data on the expression of ACE2 and TMPRSS2 in the human endometrium are sparse. Moreover, it remains unclear whether SARS-CoV-2 directly affects the embryo and its implantation. A study of the effect of SARS-CoV-2 on pregnancy showed an increase in preterm delivery. Thus, vertical transmission of the virus from mother to fetus in the third trimester is possible, and further data on human reproduction are required to establish this possibility. Based on analyses of existing data, major organizations in this field have published guidelines on the treatment of infertility. Regarding these guidelines, despite the COVID-19 pandemic, reproductive treatment is crucial for the well-being of society and must be continued under suitable regulations and good standard laboratory practice protocols.

• BMB Reports
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• 제51권12호
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• pp.648-653
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• 2018

Serine protease inhibitor Kazal type 1 (SPINK1) plays a role in protecting the pancreas against premature activation of trypsinogen and is involved in cancer progression. SPINK1 promoted LAC cells growth, migration, and invasion. Mechanistically, we found that SPINK1 promoted LAC cells migration and invasion via up-regulating matrix metalloproteinase 12 (MMP12). We observed that SPINK1 expression was only up-regulated in lung adenocarcinoma (LAC) tissues, and was an independent prognostic factor for poor survival. Our results indicate that SPINK1 might be a potential biomarker for LAC that promotes progression by MMP12.

• IMMUNE NETWORK
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• 제21권1호
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• pp.3.1-3.16
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• 2021

Coronavirus disease 2019 caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) has been spreading worldwide since its outbreak in December 2019, and World Health Organization declared it as a pandemic on March 11, 2020. SARS-CoV-2 is highly contagious and is transmitted through airway epithelial cells as the first gateway. SARS-CoV-2 is detected by nasopharyngeal or oropharyngeal swab samples, and the viral load is significantly high in the upper respiratory tract. The host cellular receptors in airway epithelial cells, including angiotensin-converting enzyme 2 and transmembrane serine protease 2, have been identified by single-cell RNA sequencing or immunostaining. The expression levels of these molecules vary by type, function, and location of airway epithelial cells, such as ciliated cells, secretory cells, olfactory epithelial cells, and alveolar epithelial cells, as well as differ from host to host depending on age, sex, or comorbid diseases. Infected airway epithelial cells by SARS-CoV-2 in ex vivo experiments produce chemokines and cytokines to recruit inflammatory cells to target organs. Same as other viral infections, IFN signaling is a critical pathway for host defense. Various studies are underway to confirm the pathophysiological mechanisms of SARS-CoV-2 infection. Herein, we review cellular entry, host-viral interactions, immune responses to SARS-CoV-2 in airway epithelial cells. We also discuss therapeutic options related to epithelial immune reactions to SARS-CoV-2.

• 미생물학회지
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• 제54권1호
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• pp.60-68
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• 2018

남극 해양세균 Pseudoalteromonas arctica PAMC 21717로부터 저온활성 alkaline protease (Pro21717)를 부분정제하였다. Pro21717 효소 추출액은 skim milk를 포함하는 zymogram gel 상에서 약 37 kDa (낮은 활성)과 74 kDa (높은 활성) 위치에서 두 개의 뚜렷한 투명밴드(clear zone)를 형성하였다. 단백질 분해활성을 나타내는 두 개의 효소단백질은 동일한 N-말단 아미노산 서열을 가지고 있었으며, 하나의 유전자에서 발현된 미성숙 단백질(precursor)이 37 kDa 크기의 단백질분해효소로 성숙화과정을 거친 후 74 kDa 크기로 이량체화됨으로써 좀 더 높은 활성을 가지는 것으로 판단된다. Pro21717은 $0-40^{\circ}C$ (최고활성 온도 $40^{\circ}C$) 온도 범위에서 단백질분해활성을 나타내었고 pH 5.0-10.0 (최적 pH 9.0) 범위에서 효소활성을 유지하였다. 주목할만한 특성으로써, Pro21717은 $40^{\circ}C$에서의 최고 효소활성(100%) 대비, $0^{\circ}C$와 $10^{\circ}C$에서 각각 30%와 45%의 높은 저온활성을 나타내었다. 또한 다양한 합성 펩타이드류에 대해 분해활성을 나타내는 Pro21717은 $Cu^{2+}$에 의해 활성이 증가하였으며, 시판용 세탁세제(commercial detergent formulation)에 포함되어 있는 다양한 종류의 계면활성제, 화학성분, 금속이온에 의해 활성이 감소되지 않았다. 전반적으로 저온활성 Pro21717은 글로벌 상업용효소 생산회사 Novozymes이 시판하고 있는 중온성 효소 Subtilisin Carlsberg (trademark Alcalase)에 버금가는 유용한 효소학적 특성이 있는 동시에 상대적으로 더 높은 저온활성을 보여주고 있다. 위의 실험결과들은, Pro21717은 $15^{\circ}C$ 이하의 차가운 수돗물에서도 세척력을 유지하는 새로운 세탁세제 효소첨가제로서의 개발 가능성을 보여주고 있다.

• 대한약리학회지
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• 제24권1호
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• pp.111-123
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• 1988

사람의 혈액으로부터 Elastase를 분리하는 새로운 방법을 개발하여 순도 높은 효소를 얻고 이 효소에 의하여 발생되어지는 질병의 예방과 치료에 응용하기 위하여 이 효소의 근본적인 성질규명을 시도하였다. 정제 방법으로는 Sephodex G-75를 이용한 1회의 액체 크로마토그라피와 HPLC을 이용한 2회의 코마토그라피를 거치는 2단계 방법을 사용하였다. 이때 얻어진 효소는 분자량이 $26,000{\sim}29,700$ 사이에 있는 3개의 다른 분자량을 가진 물질로 확인되었으며, 항체 반응에서 사람의 결구내 Elastase의 특성을 나타내었다. 함유하고 있는 미량의 금속이온을 분석한 결과 정제하는 단계에 따라 Zn 이온의 효소분자에 대한비는 증가하였으며 가장 순수한 분자내의 효소분자와 Zn 이온과의 비는 1 : 2였다. 이 효소는 chelating agent에 의하여 활성도를 잃게 되었으며, 반대로 chelating agents에 의하여 활성도를 잃고 있는 반응 medium에 그 chelaing agents의 농도를 초과하는 2가 이온 즉 Zn 이온, Ca 이온, 그리고 Mn 이온을 넣으면 그 활성도는 원상복귀되나 Mg이온에 의하여는 그 활성도가 원상회복 되지 않았다. 이 모든 성질을 종합하면 사람의 Neutrophil granule에 있는 Elastase는 지금까지 알려진 바와 같이 Serine pretense임과 동시에 metalloenzyme으로 제고되어야 한다고 제안한다. 나아가서chelating agents에 의하여 이 효소의 활성도를 조정할 수 있다는 것은 이 효소에 의하여 일어나는 질병의 치료에 응용할 수 있는 가능성도 보여준다.

• 한국축산식품학회지
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• 제22권3호
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• pp.259-266
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• 2002

본 연구는 토양으로부터 egg shell membrane(ESM)을 분해하는 미생물 strain 109를 분리 동정하였으며, 분리균이 생성한 keratinase를 정제하고 그 특성을 확인하였다. Strain 109는 16S rDNA 결과 99.9%의 상동성을 가지고 Bacillus licheniformis로 확인되었고, 3% ESM을 함유한 Nitrobacter 203배지에서 B. licheniformis 109를 접종하여 1주일간 배양하였을 때 ESM의 분해율은 약 15%였다. E. licheniformis 109가 생성한 keratinase를 정제하여 SDS-PA-GE로 분자량을 측정한 결과 약 65,000 Dalton이었으며 0.1% gelatin이 함유된 SDS-PAGE에 의해 효소 활력을 확인할 수 있었다. 정제한 keratinase의 PH에 따른 활성과 안정성은 pH 9.0에서 활성이 가장 높았으며 pH 9.0이상에서 안정하였다. 또한, 5$0^{\circ}C$에서 효소활성이 가장 높았으며 온도 안정성은 2$0^{\circ}C$에서 5$0^{\circ}C$까지 안정하였고, 7$0^{\circ}C$ 이상에서는 약 50%의 활력을 상실하였다. keratinase 활성에 금속 이온이 미치는 영향은 CuCl2와 ZnCl2에 의해 약 50% 정도가 저해되었으나 FeSO4에 의해서는 1mM일 때 약 11%, 10mM일 때 약 33%가 증가하였다. 그리고 PMSF에 의해 효소활성이 저해되는 것으로 나타나 B. licheniformis 109로부터 정제한 keratinase는 serine-protease로 사료된다.