• Journal of Microbiology and Biotechnology
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• 제23권6호
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• pp.872-877
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• 2013

In a previous study, we isolated and reported a second species of the Saccharophagus genus, Saccharophagus sp. strain Myt-1. In the present study, an alginate lyase gene (algMytC) from the genomic DNA of Myt-1 was cloned and characterized. The DNA sequence fragment obtained contained an open reading frame of 1,032 bp that encoded a protein of 343 amino acids with an estimated molecular mass of 37.6 kDa and a pI of 6.60. The deduced protein, AlgMytC, had the conserved amino acid sequences (RTELREM, QIH, YFKAGVYNQ) of the polysaccharide lyase family 7. A BLAST homology search indicated that AlgMytC shared an amino acid sequence identity of 95.9% with alg7A of S. degradans 2-40. The cloned and purified AlgMytC protein showed optimal activity at $40^{\circ}C$, and retained more than 90% of its total activity even after treatment at $25^{\circ}C$ for 24 h. AlgMytC was very alkaliphilic with an optimal pH of 9.0, and more than 90% of its activity was retained in the pH range 8.5-10.0. Moreover, AlgMytC was stable over a wide pH range. The activity of AlgMytC was also stable in the presence of various detergents.

• 생명과학회지
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• 제18권1호
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• pp.114-119
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• 2008

Wee1 인산화효소는 세포주기 조절의 핵심 단백질인 cdc2/cyclinB 복합체를 인산화하여 활성을 억제, 조절하는 주요한 효소이다. 지금까지 포유동물에서는 Wee1A, Wee1B 그리고 Myt1의 세 가지 효소가 발견되었다. Wee1 인산화효소의 조절기작을 연구하기 위하여 생쥐의 Wee1A와 Wee1B를 발톱개구리의 난자에 주사한 후 단백질의 변형을 관찰하였다. 이 세포주기 과정에서 두 효소는 모두 인산화 되었으며, Wee1A단백질은 분해되는 것을 관찰 할 수 있었다. 또한 세포외 인산화 방법을 통하여 Wee1A가 PKA와 Akt에 의해 인산화됨을 확인하였다. 이러한 Wee1 인산화효소의 인산화와 단백질 안정성에 영향을 미치는 단백질 내의 부위를 살펴보고자, Wee1A와 Wee1B의 아미노 도메인과 카르복실 도메인을 서로 치환한 단백질을 제조하여 개구리 난자에 주사하고 인산화 정도와 단백질의 안정성을 조사하였을 때, Wee1A의 아미노 도메인이 단백질의 인산화와 안정화에 중요한 영향을 미친다는 것을 규명하였다. 그리고 Wee1B의 아미노 도메인과 Wee1A의 카르복실 도메인은 효소의 활성을 조절하는 역할을 한다.

• Molecules and Cells
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• 제39권9호
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• pp.699-704
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• 2016

Developmentally regulated GTP-binding protein 2 (DRG2) plays an important role in cell growth. Here we explored the linkage between DRG2 and G2/M phase checkpoint function in cell cycle progression. We observed that knockdown of DRG2 in HeLa cells affected growth in a wound-healing assay, and tumorigenicity in nude mice xenografts. Flow cytometry assays and [$^3H$] incorporation assays indicated that G2/M phase arrest was responsible for the decreased proliferation of these cells. Knockdown of DRG2 elicited down-regulation of the major mitotic promoting factor, the cyclin B1/Cdk1 complex, but upregulation of the cell cycle arresting proteins, Wee1, Myt1, and p21. These findings identify a novel role of DRG2 in G2/M progression.

• BMB Reports
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• 제57권8호
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• pp.363-368
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• 2024

Parkinson's disease (PD), characterized by dopaminergic neuron degeneration in the substantia nigra, is caused by various genetic and environmental factors. Current treatment methods are medication and surgery; however, a primary therapy has not yet been proposed. In this study, we aimed to develop a new treatment for PD that induces direct reprogramming of dopaminergic neurons (iDAN). Achaete-scute family bHLH transcription factor 1 (ASCL1) is a primary factor that initiates and regulates central nervous system development and induces neurogenesis. In addition, it interacts with BRN2 and MYT1L, which are crucial transcription factors for the direct conversion of fibroblasts into neurons. Overexpression of ASCL1 along with the transcription factors NURR1 and LMX1A can directly reprogram iDANs. Using a retrovirus, GFP-tagged ASCL1 was overexpressed in astrocytes. One week of culture in iDAN convertsion medium reprogrammed the astrocytes into iDANs. After 7 days of differentiation, TH+/TUJ1+ cells emerged. After 2 weeks, the number of mature TH+/TUJ1+ dopaminergic neurons increased. Only ventral midbrain (VM) astrocytes exhibited these results, not cortical astrocytes. Thus, VM astrocytes can undergo direct iDAN reprogramming with ASCL1 alone, in the absence of transcription factors that stimulate dopaminergic neurons development.