• Journal of Microbiology and Biotechnology
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• 제23권9호
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• pp.1197-1205
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• 2013

Urokinase (uPA) and its receptor (uPAR) play an important role in tumor growth and metastasis. Targeting the excessive activation of this system as well as the proliferation of the tumor vascular endothelial cell would be expected to prevent tumor neovasculature and halt the tumor development. In this regard, the amino-terminal fragment (ATF) of urokinase has been confirmed as effective to inhibit the proliferation, migration, and invasiveness of cancer cells via interrupting the interaction of uPA and uPAR. Previous studies indicated that ATF expressed in Escherichia coli was mainly contained in inclusion bodies and also lacked posttranslational modifications. In this study, the biologically active and soluble ATF was cloned and expressed in Pichia pastoris. The recombinant protein was purified to be homogenous and confirmed to be biologically active. The yield of the active ATF was about 30 mg/l of the P. pastoris culture medium. The recombinant ATF (rATF) could efficiently inhibit angiogenesis, endothelial cell migration, and tumor cell invasion in vitro. Furthermore, it could inhibit in vivo xenograft tumor growth and prolong the survival of tumor-bearing mice significantly by competing with uPA for binding to cell surfaces. Therefore, P. pastoris is a highly efficient and cost-effective expression system for large-scale production of biologically active rATFs for potential therapeutic application.

• Molecules and Cells
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• 제38권5호
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• pp.402-408
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• 2015

Protein O-GlcNAcylation, dictated by cellular UDP-N-acetylglucosamine (UDP-GlcNAc) levels, plays a crucial role in posttranslational modifications. The enzyme GlcNAc kinase (NAGK, E.C. 2.7.1.59) catalyzes the formation of GlcNAc-6-phosphate, which is a major substrate for the biosynthesis of UDP-GlcNAc. Recent studies have revealed the expression of NAGK in different types of cells especially in neuronal dendrites. Here, by immunocytochemistry (ICC) and immunonucleochemistry (INC) of cultured rat hippocampal neurons, HEK293T and GT1-7 cells, we have showed that NAGK immuno-reactive punctae being present in the nucleoplasm colocalized with small nuclear ribonucleoprotein-associated protein N (snRNPN) and p54NRB, which are speckle and paraspeckle markers, respectively. Furthermore, NAGK IR cluster was also found to be colocalized with GTF2H5 (general transcription factor IIH, polypeptide 5) immuno reactive punctae. In addition, relative localization to the ring of nuclear lamin matrix and to GlcNAc, which is highly enriched in nuclear pore complexes, showed that NAGK surrounds the nucleus at the cytoplasmic face of the nuclear outer membrane. By in situ proximity ligation assay (PLA) we confirmed the colocalization of NAGK with snRNPN in the nucleus and in dendrites, while we also verified the interactions of NAGK with p54NRB, and with GTF2H5 in the nucleus. These associations between NAGK with speckle, paraspeckle and general transcription factor suggest its regulatory roles in gene expression.

• Clinical and Experimental Pediatrics
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• 제65권4호
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• pp.172-181
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• 2022

Pubertal onset is known to result from reactivation of the hypothalamic-pituitary-gonadal (HPG) axis, which is controlled by complex interactions of genetic and nongenetic factors. Most cases of precocious puberty (PP) are diagnosed as central PP (CPP), defined as premature activation of the HPG axis. The cause of CPP in most girls is not identifiable and, thus, referred to as idiopathic CPP (ICPP), whereas boys are more likely to have an organic lesion in the brain. ICPP has a genetic background, as supported by studies showing that maternal age at menarche is associated with pubertal timing in their offspring. A gain of expression in the kisspeptin gene (KISS1), gain-of-function mutation in the kisspeptin receptor gene (KISS1R), loss-of-function mutation in makorin ring finger protein 3 (MKRN3), and loss-of-function mutations in the delta-like homolog 1 gene (DLK1) have been associated with ICPP. Other genes, such as gamma-aminobutyric acid receptor subunit alpha-1 (GABRA1), lin-28 homolog B (LIN28B), neuropeptide Y (NPYR), tachykinin 3 (TAC3), and tachykinin receptor 3 (TACR3), have been implicated in the progression of ICPP, although their relationships require elucidation. Environmental and socioeconomic factors may also be correlated with ICPP. In the progression of CPP, epigenetic factors such as DNA methylation, histone posttranslational modifications, and non-coding ribonucleic acids may mediate the relationship between genetic and environmental factors. CPP is correlated with short- and long-term adverse health outcomes, which forms the rationale for research focusing on understanding its genetic and nongenetic factors.

• 한국발생생물학회지:발생과생식
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• 제13권4호
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• pp.207-215
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• 2009

시상하부-뇌하수체-생식소(hypothalamus-pituitary-gonad, HPG) 호르몬 축의 활성에 영향을 미치는 수많은 인자들은 생식 기능을 조절하고, 사춘기 개시와 폐경기 진입과 같은 뚜렷한 생식 능력의 단계 전이를 초래한다. 지방세포로부터 분비되는 다기능적 호르몬인 leptin의 발견 이후, 곧 이어 생식과 신체의 에너지 균형 사이의 긴밀한 관계에 대한 증거들이 밝혀졌다. 위장관으로부터 분비되는 또 다른 다기능 호르몬인 ghrelin은 이미 알려져 있던 growth hormone secretagogue receptor(GHSR)의 내인성 리간드이며, 에너지 항상성의 조절에서 leptin에 상응하는 물질로 알려졌다. 예상대로, ghrelin 또한 HPG 축의 활성의 조절을 통해 생식 능력을 조절함이 증명되었다. 이 논문은 ghrelin의 발견과 유전자 구조, 조직 내의 분포, 그리고 역할과 HPG 축에서의 생식 호르몬 분비 조절에 대한 포유동물의 생식에서의 ghrelin-GHSR 신호에 관한 최신 정보를 요약한 것이다. 뇌하수체에서의 POMC 유전자 발현과 유사하게, preproghrelin 유전자는 alternative splicing과 번역 후 변형(posttranslational modification)을 거치는 복잡한 레퍼토리의 전사체들과 펩티드 산물을 만들어 낸다. 에너지 항상성을 제외한 신체 생리 기능의 조절에서의 preproghrelin 유전자 산물의 역할에 관한 정보는 제한적이지만, 신진 대사와 생식 사이에서의 ghrelin의 상호작용에 관해서는 충분한 증거들이 있다. 흰쥐와 인간에서, ghrelin 수용체인 GHSRs(GHSR1a와 GHSR1b)의 분포는 본래 ghrelin의 표적으로 여겨진 시상하부와 뇌하수체뿐만 아니라 정소와 난소에서도 확인되었다. 뇌와 생식소에서도 preproghrelin 유전자 발현이 확인되었는데, 이것은 HPG 축에서 ghrelin이 국부적인 역할을 담당할 가능성을 시사한다. 비록 뇌하수체에서의 기능은 아직 확실치 않지만, ghrelin은 시상하부의 GnRH, 뇌하수체의 생식소자극호르몬과 생식소의 성 스테로이드 호르몬 분비에 대한 음성적인 조절자로서의 역할을 수행하는 것으로 보인다. 최근의 연구들은 사춘기 개시, 그리고 아마도 폐경기 진입의 조절에서 ghrelin의 관여를 시사한다. 이제 ghrelin이 '뇌-위장관' 축의 필수적인 호르몬 요인이며, 신 진 대사와 생식 사이를 연결하는 조절 물질이라는 가능성은 매우 높다. '배부름'을 반영하는 leptin 신호와는 정반대인 ghrelin 신호는 신체 에너지 균형 상태로 볼 때 '배고픔'을 표현하는 것으로 생각되며, 항상성의 유지에서 최우선 사항으로 고려되지 않는 생식으로의 에너지 투자가 이루어지지 않도록 하는데 필수적일 것으로 사료된다. 생식능력 조절에 있어서 ghrelin의 보다 명확한 작용 메커니즘과 역할에 대한 깊은 통찰력을 얻고 성공적인 생의학적 적용을 위해서는 향후 더 많은 연구들이 필요하다.

• 대한소아치과학회지
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• 제44권2호
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• pp.164-169
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• 2017

상아질 형성부전증 제 II 형은 상아질 기질에 영향을 미치는 유전질환으로 dentin sialophosphoprotien(DSPP)의 돌연변이에 기인한다. DSPP 발현 산물 중 하나인 dentin phosphoprotein(DPP)은 Asp-Ser-Ser 서열이 반복되는 독특한 구조로, 상아질 석회화 성숙 과정에 관여한다. 본 연구는 DPP 영역을 포함한 DSPP 유전자 염기 서열 분석을 통하여 유전성 상아질 결함의 원인이 되는 돌연변이를 확인하고자 하였다. 임상적 및 방사선학적 검사와 DSPP 유전자 염기 서열 분석 및 DPP 영역의 allele-specific cloning을 시행하여 비교 분석한 결과, 2688번 염기인 티민의 결실(c.2688delT)이 확인 되었고, -1 bp frameshift 돌연변이를 야기하였다. 이는 친수성 아미노산을 소수성으로 치환시켜, DPP 특성의 변화 및 상호작용 능력 저하를 일으켜 상아질 형성부전증 제 II 형의 원인으로 작용하였다.

• Asian-Australasian Journal of Animal Sciences
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• 제19권6호
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• pp.779-783
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• 2006

The adiponectin gene is known to be involved in the regulation of energy homeostasis involving food intake, carbohydrate and lipid metabolism. Human adiponectin gene polymorphisms have been recently reported to be associated with obesity, insulin sensitivity and the risk of type 2 diabetes. The present study was carried out to investigate the porcine adiponectin gene as a candidate gene for fat deposition and carcass traits. A mutation of A178G of the porcine adiponectin gene that resulted in substitution of the amino acid Isoleucine to Valine was identified. AcyI PCR-RFLP was used to detect the polymorphism of the genotypes in five different pig populations (Large White, Landrace, Duroc, Chinese breeds Meishan and Qingping). The A allele frequency was significantly higher among subjects from Chinsese lard type breeds, while the G allele was the only one present in those from Western lean type breeds. To determine if there was an association of the polymorphism with phenotypic variation, the mutation was tested in 267 pigs of the "Large $White{\times}Meishan$" F2 resource population. The results of association analyses showed significant associations of the genotypes with fat deposition and carcass traits. Allele G was significantly associated with increase in loin eye height, loin eye area and lean meat percentage and bone percentage, and decrease in fat mean percentage, ratio of lean to fat, shoulder fat thickness, 6-7 rib fat thickness, thorax-waist fat thickness and buttock fat thickness. The substitution of A178G (Ile60Val) happened to be located at amino acid 60 in the collagenous domain of porcine adiponectin which might affect the association into higher-order structures, and accordingly affect the posttranslational modifications and optimal biological activity of the multimeric forms. The identified functional polymorphism provides new evidence of adiponectin as an important candidate gene affecting fat deposition and carcass traits in pigs.