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단백질의 수용성, 열안정성 개량 기술

■ 효소의 수용성, 열안정성, 단백질 분해 효소 안정성 및 수용체 친화도 개선을 위한 초고속 탐색 기술을 개발하고 있습니다. 형광의 변화를 통하여 효소의 종류와 상관없이 모든 종류의 단백질의 수용성 (또는 상기의 성질) 변화를 측정할 수 있는 방법을 개발하고 있습니다. 이러한 기술을 이용하여 모유 올리고당인 fucosyllactose 생산 효소의 성능을 개선하는 연구 또한 수행하고 있습니다.

  그림 4. 세포 내 단백질의 ligand 결합 또는 membrane protein의 안정성을 FACS로 선별하는 HTSS법. A. ligand가 목적 단백질에 부착되어 thermostability가 높아지면 열처리에 의해 침전이 덜 일어난다. 이 때 수용성 단백질을 His6 특이적 형광염료(Ni-NTA-Ac) 등을 이용하여 검출하면 FACS를 이용한 HTSS 구현이 가능하다. B. endosome display를 이용한 membrane protein의 안정화 HTSS법.

 

수용성 단백질의 solubility, thermostability, ligand (또는 substrate) affinity 증대를 형광을 이용하여 단일세포 수준에서 측정하는 기술의 개발 및 이를 이용한 단일세포 선별법은 많은 양의 단백질을 선별하기 위해 단백질의 thermostability와 substrate specificity를 측정하는 기술로써 세포 내에서 형광을 이용하여 단일세포를 관찰하는 방법에 대해 연구하고 있습니다. 세포 내에서의 형광은 flow cytometry를 이용하여 한 번에 수십만~수백만 개의 mutant들을 테스트 할 수 있으며, 96-well micro plate를 이용하여 수천 개의 변이체 단백질들을 검증하여 후보 단백질 DNA를 얻어 낼 수 있습니다. 이는 세포 투과가 가능하며 ligand와 결합하는 단백질과 특이적으로 결합하는 표식과 형광을 내는 fluorescence dye를 합성하여 진행 하고 있습니다. 또한 이를 membrane protein의 endosome display에 응용한다면 세포 내에서 안정적으로 발현되는 membrane protein의 variation을 찾을 수 있고, 타겟 membrane protein이 endosome에 display 되어 있을 때, 이와 결합하는 파트너 단백질의 variation library를 FACS을 통하여 탐색함으로써 개선된 결합 파트너 단백질을 얻어 내는 방법을 통해 막 단백질의 protein-protein interaction 분자 진화 도구로 이용할 수 있게 하여 막 단백질의 안정화 HTSS(high throughput screening or selection) 및 target membrane protein과binding affinity이 개선된 파트너 단백질의 분자 진화 형광 탐색법으로 사용할 것입니다.

 

 

바이러스의 멤브레인에 구멍을 뚫는 나노퍼포레이터

■ 바이러스의 envelope membrane에 구멍을 뚫어 바이러스를 물리적으로 사멸하는 항바이러스제를 개발하고 있습니다.

인플루엔자 바이러스는 질병과 사망을 일으키는 주요 원인 바이러스로, 여전히 인류가 정복하지 못한 질병 중 하나입니다. 특히나 매년 나타나는 변종으로 인해 타미플루를 포함한 현재 시판된 항인플루엔자제의 경우, 그 사용에 있어 많은 제약이 따릅니다. 저희 연구실은 바이러스의 막에 구멍을 뚫어 물리적으로 사멸시키는 항바이러스제를 연구 및 개발하고 있습니다. 직경이 약 15 nm 크기로 인지질로 이루어진 이 나노 구조체는 낮은 pH 환경에서 바이러스의 막과의 융합이 가능한 구조를 지닙니다. 뿐만 아니라 원하는 수용체로 호환이 가능하다는 특징으로 인해 변종을 포함한 여러 인플루엔자 바이러스에 적용 가능한 나노 구조체 개발을 목표로 하고 있습니다.

Endocytosis하는 대장균 및 이를 이용한 생물전환 기술 

■ 대장균이 endocytosis 할 수 있도록 engineering 함으로써 막단백질의 대량 생산, 막단백질의 engineering 기술의 개발, 지방산의 생물 전환 등에 획기적인 전환점을 만들고 있습니다. 

  Caveolae 는 척추동물 내에서 막 굴곡 및 세포 내 소포를 만드는 구조 중 하나입니다. 최근 대장균 내에서 Caveolae 구조를 모사할 수 있다는 연구가 보고 되었습니다. 본 실험실에서는 이 Caveolae 형성 대장균을 여러 분야에 활용하고자 합니다. 첫 번째로, 불용성 막단백질을 Caveolae 에 함께 발현시킴으로써, 막단백질의 연구에 응용하고자 합니다. 기존의 방법을 이용하여 막단백질을 연구하려면, 막단백질의 발현 및 정제, 그리고 인공의 막에 삽입 등 여러 가지 과정들이 필요합니다. 여러 과정 중에서 막단백질은 종종 Inclusion body 등을 형성하여, 정제 효율이 떨어지거나, 활성을 잃기도 합니다. 반면에 Caveolae 에 막 단백질을 삽입하게 되면, Caveolae 에 막 단백질이 Display 된 구조가 만들어지게 되어, Inclusion body의 형성을 막아주고, Caveolae 위에서 막단백질 본래의 활성을 유지하게 됩니다. 두 번째로, 대장균 cytosol 내부에 지질친화적 물질을 수송할 때 Caveolae를 이용하고자 합니다. 대장균을 이용한 지질친화적인 지방산의 전세포 반응 등을 진행하기 위해선, 지방산을 대장균의 cytosol로 수송해야 합니다. 이 때, 지방산은 대장균의 외막 내막을 통과해야 하는데, Caveolae 를 만들어 세포 내 함입이 일어나는 대장균은 지방산의 수송이 향상됨을 확인하였습니다.

 

 

SNARE가 일으키는 매우 빠른 막융합

■ SNARE 단백질이 어떻게 칼슘에 의해서 엄청나게 빠른 속도로 membrane fusion을 일으킬 수 있는지에 대한 의문에 대해 공부하고 있습니다. 이러한 호기심을 풀어가는 과정에서 식물성의 저분자 화합물을 이용하여 보톡스(주름 제거 등에 널리 이용되는 신경독)을 대체하는 기술을 개발하고, 현재 이의 산업과 연구를 수행하고 있습니다.

 

보툴리눔 신경독(일명, 보톡스)은 SNARE라 불리는 막융합 단백질을 절단하여 신경 전달 과정을 조절함으로써 주름 제거 등 다양한 효능을 보이는 의약품입니다. 본 연구실은 미리세틴이라고 하는 천연 저분자 플라보노이드는 SNARE 단백질의 복합체 형성 과정에 관여하여 보톡스와 유사한 효능을 나타낸다는 것을 밝힌 적이 있어서 이를 저분자 식물성 보톡스라 이름 붙인 적이 있습니다.

최근에는 이를 이용하여 엄청나게 빠른 신경전달의 조절 기전을 밝힐 수 있었습니다. 연구의 결과는 Journal of American Chemical Society (Impact factor, 12.113) 2016년 3월호에 발표되었습니다. 아래에 그 내용을 간략히 정리하였습니다.

1) 미리세틴은 신경소낭과 신경세포막을 반융합 상태에서 정지시킨다.

 

-   스네어 단백질은 아미노말단에서 카르복실말단까지 지퍼가 채워지듯이 헬릭스 다발을 만드는데, 이때 미리세틴이 스네어 단백질의 지퍼 채우는 과정의 중간에 끼어들어서 지퍼가 더 이상 채워지지 않게 만듬.

-   결과적으로 막융합이 반만 일어난 반융합 상태에서 정지됨. 이는 신경전달이 차단됨을 의미함.

-   보톡스는 이 스네어 단백질을 절단하는 단백질 분해 효소임에 반해 미리세틴은 스네어 단백질의 지퍼링을 저해한다는 측면에서 차이가 있음.

 

2) 반융합 상태에서 정지된 스네어 지퍼는 미리세틴을 빼내면 막융합 반응을 재개할 수 있다.

-    락케이즈라고 하는 효소로 스네어 지퍼에 끼어 있는 미리세틴을 빼내면, 스네어는 지퍼를 끝까지 채울 수 있고, 반융합 상태에 머물러 있던 막융합도 완성될 수 있다.

-    시냅토태그민이라고 하는 칼슘 센서 단백질이 있으면 칼슘 유입과 거의 동시에 최종의 막융합을 일으킨다.

 

3) 신경소낭이 반유합 상태에 있을 때 매우 빠르고 잘 동기화된 신경전달이 가능하다.

 

-    반융합이 이루어지지 않은 상태에서 칼슘이 유입되면 수초에 걸쳐서 천천히 막융합이 일어난다.

-    반융합 상태에서 칼슘이 유입되면 거의 모든 신경소낭의 막융합 반응이 동시에 수 밀리초 이내에 일어난다. 이는 신경 전달이 매우 빠르게 일어나기 위해서 반융합 상태에서 일시적으로 막융합을 정지시키는 것이 매우 중요함을 의미한다.

천연물을 이용한 보톡스 대체 기술 개발

빠른 신경전달 과정에 관한 수 십년 간의 미스터리를 해결하였을 뿐만 아니라 약물을 이용하여 신경전달 과정을 조절하는 것이 가능함을 보였습니다. 저분자 천연물을 이용하여 보톡스를 대체할 수 있는 기술에 대한 과학적 근거를 마련하였다고 생각합니다. 보톡스는 매우 치명적인 신경독이어서 전문의의 시술을 통해서만 사용 가능한데, 저분자 천연물 보톡스는 화장품 등 다양한 형태로 개발이 가능합니다. 보톡스는 주름 제거 뿐만 아니라 다한증, 요실금 등 매우 다양한 적응증에 사용되는 등 광범위한 치료 효능을 가지는데 (세계 시장 규모 4조원), 저분자 천연물 보톡스는 기존의 보톡스를 일정 부분 대체할 수 있을 뿐만 아니라 보톡스가 적용될 수 없는 다양한 상품 및 치료제의 개발에 응용될 수 있습니다.

그러나 이런 천연 유래의 저분자 물질들은 색이 강하고 외부 자극에 불안정하며 용해도가 낮아 화장품 및 의야 소재로 사용이 어렵습니다. 저희 연구실에서는 이러한 단점을 극복하기 위하여 화학적/생화학적 반응을 통해 천연 저분자 물질의 안정성 및 세포 투과성을 향상 시키는 연구 또한 수행하고 있습니다. 또한, 저분자 보톡스 효능을 내는 천연물을 추가적으로 탐색하고 있습니다.

 

관련 논문

-       Paul Heo, Byoungjae Kong, Younghoon Jung, Joon-Bum Park, Jong-Hyeok Shin, Myeongseo Park, Dae-Hyuk Kweon. Green fluorescence protein-based content-mixing assay of SNARE-driven membrane fusion. Biochemical and Biophysical Research Communications, 2017, 488, 53-59.

-       Paul Heo, Joon-Bum Park, Yeon-Kyun Shin and Dae-Hyuk Kweon. Visualization of SNARE-mediated hemifusion between giant unilamellar vesicles arrested by myricetin. Frontiers in Molecular Neuroscience, 2017, 10, Article 92, 1-10.

-       Dae-Hyuk Kweon, Byoungjae Kong and Yeon-Kyun Shin. Hemifusion in synaptic vesicle cycle, Frontiers in Molecular Neuroscience, 2017, 10, Article 65, 1-8.

-       Tae-Sun Lee, Joo-Young Lee, Jae Won Kyung, Yoosoo Yang, Seung Ju Park, Seulgi Lee, Igor Pavlovic, Byoungjae Kong, Yong Seok Jho, Henning J Jessen, Dae-Hyuk Kweon, Yeon-Kyun Shin, Sung Hyun Kim, Tae-Young Yoon, Seyun Kim. Inositol Pyrophosphates Inhibit Synaptotagmin-Dependent Exocytosis, Proceedings of the National of Academy of Sciences of the USA, 2016, 113(29) 8314-8319.

-       Paul Heo, Yoosoo Yang, Kyu-Young Han, Byoungjae Kong, Jong-Hyeok Shin, Younghoon Jung, Cherlhyun Jeong, Jaeil Shin, Yeon-Kyun Shin, Taekjip Ha and Dae-Hyuk Kweon. A chemical controller of SNARE-driven membrane fusion that primes vesicles for Ca2+-triggered millisecond exocytosis, Journal of the American Chemical Society, 2016, 138 (13), pp 4512–4521

-       Yoosoo Yang; Paul Heo; Byoungjae Kong; Jun-Bum Park; Young-Hun Jung; Jonghyeok Shin; Cheol-Hyun Jung, Dae-Hyuk Kweon (2015) Dynamic light scattering analysis of SNARE-driven membrane fusion and the effects of SNARE-binding flavonoids, Biochemical and Biophysical Research Communications, 465, 864-870.

-       Yoosoo Yang, Se-Hyun Kim, Paul Heo, Byoungjae Kong, Jonghyeok Shin, Young-Hun Jung, Keejung Yoon, Woo-Jae Chung, Yeon-Kyun Shin, Dae-Hyuk Kweon. (2014) SNARE zippering is hindered by polyphenols in the neuron. Biochemical and Biophysical Research Communications, 450, 831-836.

-       Jaeil Shin, Dae-Hyuk Kweon, Yeon-Kyun Shin. (2014) Multiple conformations of a single SNAREpin between two nanodisc membranes reveals diverse pre-fusion states. Biochemical Journal, 459, 95–102.

-       Bong-Kyu Choi, Mal-Gi Choi , Jae-Yeol Kim , Yoosoo Yang , Ying Lai , Dae-Hyuk Kweon , Nam Ki Lee , Yeon-Kyun Shin (2013) Large α-Synuclein oligomers inhibit neuronal SNARE-mediated vesicle docking. Proceedings of the National of Academy of Sciences of the USA, 110 (10) 4087-4092.

-       Yoosoo Yang, Jung-Mi Oh, Paul Heo, Jae Yoon Shin, Byoungjae Kong, Jonghyeok Shin, Ji-Chun Lee, Jeong Su Oh, Kye Won Park, Chung Hwan Lee, Yeon-Kyun Shin, Dae-Hyuk Kweon (2013) Polyphenols differentially inhibit degranulation of distinct subsets of vesicles in mast cells by specific interaction with granule type-dependent SNARE complexes. Biochemical Journal, 450 (3), 537-546.

-       Chang Hwa Jung, Jin Kyu Choi, Yoosoo Yang, Hyun Ju Koh, Paul Heo, Kee-Jung Yoon, Sehyun Kim, Won-Seok Park, Hong-Ju Shin, Dae-Hyuk Kweon. A botulinum neurotoxin-like function of Potentilla chinensis extract that inhibits neuronal SNARE complex formation, membrane fusion, neuroexocytosis, and muscle contraction, (2012) Pharmaceutical Biology, 50(9) 1157-1167.

-       Yoosoo Yang, Jin Kyu Choi, Chang Hwa Jung, Hyun Ju Koh, Paul Heo, Jae Yoon Shin, Sehyun Kim, Won-Seok Park, Hong-Ju Shin, Dae-Hyuk Kweon (2012) SNARE-wedging polyphenols as small molecular Botox, Planta Medica, 78(3) 233-236.

-       Tae-Young Yoon, Dae-Hyuk Kweon, Yeon-Kyun Shin (2011) Chasing the trails of SNAREs and lipids along the membrane fusion pathway. Current Topics in Membranes. 68, 161-184.

-       Jun-Seob Kim, Paul Heo, Tae-Jun Yang, Ki-Sung Lee, Yong-Su Jin, Sung-Koo Kim, Dongwoo Shin and Dae-Hyuk Kweon (2011) Bacterial persisters tolerate antibiotics by not producing hydroxyl radicals. Biochemical and Biophysical Research Communications 413, 105-110.

-       Yoo-Soo Yang, Jae Yoon Shin, Jung-Mi Oh, Chang Hwa Jung, Yunha Hwang, Se-Hyun Kim, Jun-Seob Kim, Kee-Jung Yoon , Ji-Young Ryu, Jaeil Shin, Jae Sung Hwang, Tae-Young Yoon, Yeon-Kyun Shin and Dae-Hyuk Kweon (2010) Dissection of SNARE-driven membrane fusion and neuroexocytosis by wedging small hydrophobic molecules into the SNARE zipper, Proceedings of the National of Academy of Sciences of the USA, 107(51) 22145-22150.

-       Han-Ki Lee, Yoosoo Yang, Zengliu Su, Changbong Hyeon, Tae-Sun Lee, Hong-Won Lee, Dae-Hyuk Kweon, Yeon-Kyun Shin and Tae-Young Yoon (2010) Dynamic Ca2+-dependent stimulation of vesicle fusion by membrane-anchored synaptotagmin 1, Science, 328, 760-763.

-       Chang Hwa Jung, Jeong-Hyuk Kim, Sunju Park, Dae-Hyuk Kweon, Sung-Hoon Kim and Seong-Gyu Ko (2010) Inhibitory effect of Agrimonia pilosa Ledeb. On inflammation by suppression of iNOS and ROS production, Immunological Investigations, 39(2), 159-170.

-       Yoo-Soo Yang, Jae-Il Shin, Jae-Yoon Shin, Jung-Mi Oh, Sang Ho Lee, Dae-Hyuk Kweon (2009) Prion Protein Does Not Interfere with SNARE Complex Formation and Membrane Fusion, Food Science and Biotechnology, 18(3) 782-787.

-       Chang Hwa Jung, Yoo Soo Yang, Jun-Seob Kim, Yeon-Kyun Shin, Jae Sung Hwang, Eui Dong Son, Hong Hwa Lee, Koo Min Chung, Jung Mi Oh, Jong Hwa Lee, Dae-Hyuk Kweon (2009) Inhibition of SNARE-driven neuroexocytosis by plant extracts. Biotechnology Letters, 31, 361-369.

-       Chang Hwa Jung, Yoo Soo Yang, Jun Seob Kim, Jae Il Shin, Yong-Su Jin, Jae Yoon Shin Jong Hwa Lee, Koo Min Chung, Yeon-Kyun Shin, and Dae-Hyuk Kweon (2008) A Search for Synthetic Peptides that Inhibit SNARE-Mediated Membrane Fusion. FEBS Journal, 275, 3051-3063.

-       Kweon, D.-H., Kim, C. S., and Shin Y.-K. (2003) Regulation of neuronal SNARE assembly by the membrane. Nat. Struct. Biol. 10, 440-447.

-       Kweon, D.-H., Kim, C. S., and Shin Y.-K. (2003) Insertion of the membrane proximal region

of the neuronal SNARE coiled coil into the membrane. J. Biol. Chem. 278, 12367-12373.

-       Kweon, D.-H., Kim, C. S., and Shin Y.-K. (2002) The membrane-dipped neuronal SNARE complex: A site-directed spin labeling electron paramagnetic resonance study. Biochemistry 41, 9264-9268.

-       Zhang, F., Chen, Y., Kweon, D.-H., Kim, C. S., and Shin, Y.K. (2002) The four-helix bundle of the neuronal target membrane SNARE complex is neither disordered in the middle nor uncoiled at the C-terminal region. J. Biol. Chem.  277, 24294-24298.

-       Kweon, D.-H., Chen, Y., Zhang, F., Poirier M., Kim, C. S., and Shin, Y.K. (2002) Probing domain swapping for the neuronal SNARE complex with electron paramagnetic resonance. Biochemistry 41, 5449-5452.

-       Kim, C. S, Kweon, D.-H., and Shin, Y.K. (2002) Membrane topologies of neuronal SNARE folding intermediates. Biochemistry 41, 10928-10933.

 

 

재조합 미생물을 이용한 유용 물질의 생산

■ 재조합 미생물을 이용한 외래 단백질 및 유용물질의 생산

재조합 미생물(대장균 및 효모 등)를 이용하여 유용한 외래 단백질을 대량 생산하는 연구, 항생제 내성을 극복하는 방법에 관한 연구, 효소에서의 젖산 생산에 관한 연구, 효모 및 미생물의 유전공학적 기술 개발에 관한 연구 등을 수행하고 있습니다.

-       Jae Won Lee, Jung Hoon In, Jun-Bum Park, Jonghyeok Shin, Jin Hwan Park, Bong Hyun Sung, Jung-Hoon Sohn, Jin-Ho Seo, Jin-Byoung Park, Soo Rin Kim, Dae-Hyuk Kweon. Co-expression of two heterologous lactate dehydrogenases genes in Kluyveromyces marxianus for L-lactic acid production, Journal of Biotechnology, 2017, 241, 81-86.

-       Jun-Seob Kim, Da-Hyeong Cho, Paul Heo, Suk-Chae Jung, Myungseo Park, Eun Joong Oh, Jaeyun Sung, Pan-Jun Kim, Sukchan Lee, Dae-Hee Lee, Sarah Lee, Choong Hwan Lee, Dongwoo Shin, Yong-Su Jin, and Dae-Hyuk Kweon (2016) Fumarate-mediated persistence of Escherichia coli against antibiotics. Antimicrobial Agents and Chemotherapy, 60(4) 2232-2240.

-       Jun-Seob Kim, Da-Hyeong Cho, Myeongseo Park, Woo-Jae Chung, Dongwoo Shin, Kwan Soo Ko, and Dae-Hyuk Kweon (2016) CRISPR/Cas9-mediated re-sensitization of antibiotic-resistant Escherichia coli harboring extended spectrum b-lactamases. Journal of Microbiology and Biotechnology, 26(2) 394-401.

-       Hyewoo Ppyun, Seung Hyun Kim, Man Hui Youn, Sung Suk Cho, Kyung Min Kwon, Dae-Hyuk Kweon, Suk-Tae Kwon (2015) Improved PCR performance and fidelity of double mutant Neq A523R/N540R DNA polymerase. Enzyme and Microbial Technology 82, 197-204.

-       Jonghyeok Shin, Younghun Jung, Da-Hyoung Cho, Myeongseo Park, Kyungeun Lee, YoosooYang, Cherlhyun Jeong, Bong Hyun Sung, Jung-Hoon Sohn, Jin-Byoung Park, Dae-Hyuk Kweon (2015) Display of membrane proteins on the heterologous caveolae carved by caveolin-1 in the Escherichia coli cytoplasm, Enzyme and Microbial Technology, 79-80, 55-62.

-       Ji-Chun Lee, Byoung-Jae Kong, Jae Yoon Shin, Jonghyeok Shin, Jung-Su Oh, Jae Youl Cho, Keejung Yoon, Sung-Gun Kim, Young-Ha Song, Chol-Su Shin, Heekyung An, Woo-Jong Lee and Dae-Hyuk Kweon (2015) A pH-Responsive High Density Lipoprotein-Like Nanoparticle of Epothilone B, Biotechnology Bioprocess Engineering, 20(2) 249-258.

-       Kwon CW, Park KM, Kang BC, Kweon DH, Kim MD, Shin SW, Je YH, Chang PS. (2015) Cysteine Protease Profiles of the Medicinal Plant Calotropis procera R. Br. Revealed by De Novo Transcriptome Analysis. PLoS One. 2015 Mar 18;10(3):e0119328

-       Jang Jiwon, Byun Sung-Hyun, Han Dasol, Lee Junsub, Kim Juwan, Lee Nayeon, Kim Inhee, Park Soojeong, Ha Soobong, Kwon Mookwang, Ahn Jyhyun, Chung Woo-Jae, Kweon Dae-Hyuk, Cho Jae Youl, Kim Sunyoung, and Yoon Keejung. (2014) Notch Intracellular Domain Deficiency in Nuclear Localization Activity Retains the Ability to Enhance Neural Stem Cell Character and Block Neurogenesis in Mammalian Brain Development. Stem Cells and Development. 23(23) 2841-2850.

-       Muhammad Jahangir Hossen, Kwang-Soo Baek, Eunji Kim, Woo Seok Yang, Deok Jeong, Jun Ho Kim a, Dae-Hyuk Kweon, Deok Hyo Yoon, Tae Woong Kim, Jong-Hoon Kim, Jae Youl Cho (2014) In vivo and in vitro anti-inflammatory activities of Persicaria chinensis methanolic extract targeting Src/ Syk/NF-κB. Journal of Ethnopharmacology.

-       Yuna Cheon, Jun-Seob Kim, Jun-Bum Park, Paul Heo, Jae Hyung Lim, Gyoo Yeol Jung, Jin-Ho Seo, Jin Hwan Park, Hyun Min Koo, Kwang Myung Cho, Jin-Byung Park, Suk-Jin Ha, Dae-Hyuk Kweon (2015) A biosynthetic pathway for hexanoic acid production in Kluyveromyces marxianus Journal of Biotechnology 182–183 (2014) 30–36

-       Jaeil Shin, Dae-Hyuk Kweon, Yeon-Kyun Shin. (2014) Multiple conformations of a single SNAREpin between two nanodisc membranes reveals diverse pre-fusion states. Biochemical Journal, 459, 95–102.

-       Yi-Hyun Bae, Yong-Cheol Park, Dae-Hyuk Kweon, Jin-Ho Seo. (2014) Deletion of the HXK2 gene in Saccharomyces cerevisiae enables mixed sugar fermentation of glucose and galactose in oxygen-limited conditions. Process Biochemistry, 49(4), 547-553.

-       Jiwon Jang, Jyhyun Ahn, Seong-Tae Kim, Dae-Hyuk Kweon, Jae Youl Cho, Kye Won Park, Sunyoung Kim, Keejung Yoon (2013) Ultrasound backscatter microscopy image-guided intraventricular gene delivery at murine embryonic age 9.5 and 10.5 produces distinct transgene expression patterns at the adult stage. Molecular Imaging & Biology, 12(8), 1-6.

-       No-Joon Song, Hyang-Jin Yoon, Ki Hyun Kim, So-Ra Jung, Woo-Seok Jang, Cho-Rong Seo, Young Min Lee, Dae-Hyuk Kweon, Joung-Woo Hong, Jeong-Soo Lee, Ki-Moon Park, Kang Ro Lee and Kye Won Park (2013) Butein is a novel anti-adipogenic compound, Journal of Lipid Research, 54, 1385-1396.

-       Paul Heo, Tae-Jun Yang, Soon-Chun Chung, Yuna Cheon, Jun-Seob Kim, Jun-Bum Park, Hyun Min Koo, Kwang Myung Cho, Jin-Ho Seo, Jae Chan Park, Dae-Hyuk Kweon (2013) Simultaneous integration of multiple genes into the Kluyveromyces marxianus chromosome, Journal of Biotechnology 167(3) 323-325.

-       Soo Rin Kim, Ki-Sung Lee, In Iok Kong, Anastashia Lesmana, Won-Heong Lee, Jin-Ho Seo, Dae-Hyuk Kweon, Yong-Su Jin (2013) Construction of an efficient xylose-fermenting diploid Saccharomyces cerevisiae strain through mating of two engineered haploid strains capable of xylose assimilation. Journal of Biotechnology, 164 (1), 105-111.

-       Yongwoo Jung, Young-Su Yi, Dae Sung Yoo, Ji Hye Kim, Woo Seok Yang, Jongsung Lee, Gye Won Park, Dae Hyuk Kweon, Sungyoul Hong, Jae Youl Cho (2013) 8-(tosyl amino)quinoline inhibits tumour progression through targeting phosphoinositide-3-kinase/Akt pathway. Pharmazie, 68, 146–152.

-       Jae-Yoon Shin, Yoo-Soo Yang, Paul Heo, Ji-Chun Lee, Byung-Je Kong, Jae-Yeol Cho, Kee-Jung Yoon, Cheol-Su Shin, Jin-Ho Seo, Sung-Gun Kim, and Dae-Hyuk Kweon (2012) pH-responsive high density lipoprotein-like nanoparticle to release paclitaxel at acidic pH for cancer chemotherapy, International Journal of Nanomedicine, 7, 2805-2816.

-       Ki-Sung Lee, Jun-Seob Kim, Paul Heo, Tae-Jun Yang, Young-Je Sung, Yuna Cheon, Hyun Min Koo, Byung Jo Yu, Jin-Ho Seo, Yong-Su Jin, Jae Chan Park and Dae-Hyuk Kweon (2012) Characterization of Saccharomyces cerevisiae promoters for heterologous gene expression in Kluyveromyces marxianus, Applied Microbiology and Biotechnology, 97(5):2029-2041.

-       Hang, Nguyen Thi, Sung-Gun Kim, and Dae-Hyuk Kweon (2012) Electrostatic Immobilization of D-Xylose Isomerase to Cation Exchanger for Conversion of D-Xylose to D-Xylulose. Korean J. Microbiol. Biotechnol. 40(2) 163–167.

-       Jun-Seob Kim, Paul Heo, Tae-Jun Yang, Ki-Sing Lee, Da-Hyeong Cho, Bum Tae Kim, Ji-Hee Suh, Hee-Jong Lim, Dongwoo Shin, Sung-Koo Kim and Dae-Hyuk Kweon (2011) Selective killing of bacterial persisters by a single chemical compound without affecting normal antibiotic-sensitive cells. Antimicrobial Agents and Chemotherapy, 55(11) 5380-5383.

-       Tae-Young Yoon, Dae-Hyuk Kweon, Yeon-Kyun Shin (2011) Chasing the trails of SNAREs and lipids along the membrane fusion pathway. Current Topics in Membranes. 68, 161-184.

-       Jun-Seob Kim, Paul Heo, Tae-Jun Yang, Ki-Sung Lee, Yong-Su Jin, Sung-Koo Kim, Dongwoo Shin and Dae-Hyuk Kweon (2011) Bacterial persisters tolerate antibiotics by not producing hydroxyl radicals. Biochemical and Biophysical Research Communications 413, 105-110.

-       Myung-Soon Yang, Jin-Su Hong, Seong-Tae Kim, Ki-Young Lee, Kye Won Park, Suk-Tae Kwon, Dae-Hyuk Kweon, Young Ho Koh, Nicholas Gaiano and Keejung Yoon (2011) Among γ-secretase substrates Notch1 alone is sufficient to block neurogenesis but does not confer self-renewal properties to neural stem cells, Biochemical and Biophysical Research Communications, 404(1) 133-138.

-       Won-Ki Min, Dae-Hyuk Kweon, Kyungmoon Park, Yong-Cheol Park, and Jin-Ho Seo (2010) Characterization of Monoclonal Antibody against Aflatoxin B1 Produced in Hybridoma 2C12 and its Single-Chain Variable Fragment Expressed in Recombinant Escherichia coli. Food Chemistry, 126(3) 1316-1323.

-       Nam D, Choi E, Kweon D-H and Shin D (2010) The RstB sensor acts on the PhoQ sensor to control expression of PhoP-regulated genes. Molecules and Cells, 30, 363-368.

-       Kim SR, Lee KS, Choi JH, Ha SJ, Kweon DH, Seo JH and Jin YS. (2010) Repeated batch fermentation of xylose and glucose-xylose mixtures using a respiration-deficient Saccharomyces cerevisiae engineered for xylose metabolism. Journal of Biotechnology 150, 404-407.

-       Ki-Sung Lee, Dae-Hyuk Kweon and Yong-Su Jin. (2010) Improved galactose fermentation of Saccharomyces cerevisiae through inverse metabolic engineering. Biotechnology and Bioengineering, 108(3), 621-631.

-       Min-Eui Hong, Ki-Sung Lee, Byung Jo Yu, Young-Je Sung, Sung Min Park, Hyun Min Koo, Dae-Hyuk Kweon, Jae Chan Park, Yong-Su Jin (2010) Identification of gene targets eliciting improved alcohol tolerance in Saccharomyces cerevisiae through inverse metabolic engineering, Journal of Biotechnology, 149, 52-59.

-       Suk-Chae Jung, Chris L. Smith, Ki-Sung Lee, Min-Eui Hong, Dae-Hyuk Kweon, Gregory Stephanopoulos, and Yong-Su Jin  (2010) Restoration of growth phenotypes of Escherichia coli DH5 in minimal media through reversal of a point mutation in purB. Applied and Environmental Microbiology, 76, 6307-6309.

-       Chang Hwa Jung, Jeong-Hyuk Kim, Sunju Park, Dae-Hyuk Kweon, Sung-Hoon Kim and Seong-Gyu Ko (2010) Inhibitory effect of Agrimonia pilosa Ledeb. On inflammation by suppression of iNOS and ROS production, Immunological Investigations, 39(2), 159-170.

-       Ji-Young Bae, Dae-Hyuk Kweon and Joo-Sung Yang. (2009) Recombinant DNA and protein vaccines for foot-and-mouth disease induce humoral and cellular immune responses in mice. Immune Network, 9(6), 265-273.

-       Jae-Yoon Shin, Jae-Il Shin, Jun-Seob Kim, Yoo-Soo Yang, Yun-Ha Hwang, Joo-Sung Yang, Dongwoo Shin, Jin-Ho Seo, Yong-Su Jin, Yong-Cheol Park, Jae-Sung Hwang, Dae-Hyuk Kweon (2009) Assembly of Coenzyme Q10 Nanostructure Resembling Nascent Discoidal High Density Lipoprotein Particle, Biochemical Biophysical Research Communications, 388(2) 217-221.

-       Jieun Lee, Eunjong Seo, Dae-Hyuk Kweon, Kimoon Park, Yong-Su Jin (2009) Fermentation of rice bran and defatted rice bran for butanol production using Clostridium beijerinckii NCIMB 8052, Journal of Microbiology and Biotechnology 19(5) 482-490.

-       Jae Yoon Shin, Jae Il Shin, Jun Seob Kim, Yoo Soo Yang, Yeon-Kyun Shin, Kyeong Kyu Kim3, Sangho Lee, Dae-Hyuk Kweon (2009) Disulfide bond as a structural determinant of prion protein membrane insertion, Molecules and Cells, 27, 673-680.

-       Dae-Hee Lee, Sung-Gun Kim, Dae-Hyuk Kweon and Jin-Ho Seo (2009) Folding machineries displayed on a cation-exchanger for the concerted refolding of cysteine- or proline-rich proteins. BMC biotechnology, 9, article No. 27.

-       Young-Jin Son, Chang-Kyu Kim, Young Bong Kim, Dae-Hyuk Kweon, Yong-Cheol Park and Jin-Ho Seo (2009) Effects of Citraconylation on Enzymatic Modification of Human Proinsulin Using Trypsin and Carboxypeptidase B. Biotechnology Progress25(4) 1064-1070..

-       Kyungwon Lee, Jae-Yoon Shin, Yoo-Soo Yang, Jae-Il Shin, Yong-Cheol Park, Jin-Ho Seo, Tai Hyun Park, Chol-Su Shin, Yong-Su Jin, and Dae-Hyuk Kweon. (2009) Towards a microarray of functional membrane proteins: Assembly of a surface-attachable, membrane-protein-anchored membrane structure using apolipoprotein A-1. Enzyme and Microbial Technology, 44, 217-222.

-       Jaeyoung Kim, Hoyoung Chung, Suyeon Kim, Heejin Park, Dae-Hyuk Kweon (2009) Discovery of single nucleotide polymorphisms in FABP3 and leptin gene in pig. Archiv Tierzucht, 52, 106-107.

-       Ki-Sung Lee, Jae Yoon Shin, Yi-Hyun Jang, Dae-Hyuk Kweon, Kimoon Park, and Yong-Su Jin (2008) Production of casein phosphopeptides using Streptococcus faecalis var. liquefaciens cell immobilization. Korean J. Biotechnol. Bioeng. 23, 59-64.

-       Suk-Chae Jung, Junseob Kim, Chang Yong Jung, Dae-Hyuk Kweon, Kimoon Park, and Yong-Su Jin. (2007) Policosanol production from rice bran oil byproducts. Food Engineering Progress, 11, 293-297.

-       Jeong, H. J., Jeong, J. B., Kim D. S., Park, J. H., Lee, J. B. Kweon, D.-H., Chung, G. Y. Seo, E. W., and Lumen, B. O. (2007) The cancer preventive peptide lunasin from wheat inhibits core histone acetylation. Cancer Letters 255, 42-48.

-       Jang, C.-M., Kweon, D.-H., and Lee, J.-H. (2007) Effect of mucilage from yam on activation of lymphocytic immune cells. Nutrition Research and Practice. 2, 94-99.

-       Lee, J-H, Kweon, D.-H., Lee, S.-C. (2006) Isolation and characterization of an immunopotentiating factor from Lactobacillus plantarum in Kimchi: Assessment of immunostimulatory activities. Food Sci. Biotechnol. 15, 877-883.

-       Kim, S-J., Kweon, D.-H., and Lee, J. H. (2006) Investigation of antioxidative activity and     stability of ethanol extracts of licorice root (Glycyrrhiza glabra). K. J. Food Sci. Technol. 38, 584-588.

-       Kim, S.-G., Kim, J.-A., Yu, H.-A., Lee, D,-H, Kweon, D.-H., and Seo J.-H. (2006) Application of poly-arginine fused minichaperone to renaturation of cyclodextrin glycosyltransferase expressed in recombinant Escherichia coli. Enz. Microbial. Technol. 39,495-465.

-       Kim, S-J., Shin, J. Y., Park, Y. M. Chung, K. M. Lee, J. H., and Kweon, D.-H. (2006) Investigation of antimicrobial activity and stability of ethanol extracts of licorice root (Glycyrrhiza glabra). K. J. Food Sci. Technol. 38, 241-248.

-       Rha, C.-S., Lee, D.-H., Kim, S.-G., Min, W.-K., Byun, S.-G., Kweon, D.-H., Han, N. S. and Seo, J.-H. (2005) Production of cyclodextrin by poly-lysine fused Bacillus macerans cyclodextrin glycosyltransferase immobilized on cation exchanger. J. Mol. Cat. B:Enzymatic 34, 39-43,

-       Kweon, D.-H., Kim, S. G., Han, N. S. Lee, J. H., Chung, K. M. and Seo, J.-H. (2005) Immobilization of Bacillus macerans cyclodextrin glycosyltransferase fused with poly-lysine using cation exchanger. Enz. Microbial Technol. 36, 71-578.

-       Kim, S. G., Kweon, D.-H., Lee D.-H., Park, Y.-C., and Seo, J.-H. (2005) Coexpression of folding accessory proteins for production of active cyclodextrin glycosyltransferase of Bacillus macerans in recombinant Escherichia coli. Prot. Exp. & Puri. 41, 426-432.

-       Kweon, D.-H., Kim S. G., and Seo, J.-H. (2004) Purification and Refolding of Cyclodextrin

-       Glycosyltransferase Expressed from Recombinant Escherichia coli J. Incln Pheno. 50, 37-41.

-       Choi, G.-H, Lee, D.-H., Min, W.-K., Cho, Y.-J., Kweon, D.-H., Son, D.-H., Park, K.-M. and Seo, J.- H. (2004) Cloning, expression, and characterization of single-chain variable fragment (scFv) antibody against mycotoxin deoxynivalenol in recombinant Escherichia coli. Prot. Exp. & Puri. 35, 84-92.

-       Lee, D.-H., Kweon, D.-H., Han, N. S., Seo, J.-H. (2004) Consortium of fold-catalyzing proteins increases soluble expression of cyclohexanone monooxygenase in recombinant Escherichia coli. Appl. Microbiol. Biochnol. 63, 549-552.

-       Kim, H.J., Lee D.-Y., Lee, D.-H., Park, Y.-C., Kweon, D.-H., Ryu, Y.-W. and Seo, J.-H. (2004) Strategic proteome analysis of Candida magnoliae with an unsequenced genome. Proteomics 4, 3588-3599.

-       Kweon, D.-H., Lee, D.-H., Han, N. S., and Seo, J.-H. (2003) Solid-state refolding of cyclodextrin glycosyltransferase. Biotechnol. Prog..20, 277-283.

-       Kweon, D.-H., Kim, C. S., and Shin Y.-K. (2003) Regulation of neuronal SNARE assembly by the membrane. Nat. Struct. Biol. 10, 440-447.

-       Kweon, D.-H., Kim, C. S., and Shin Y.-K. (2003) Insertion of the membrane proximal region

-       of the neuronal SNARE coiled coil into the membrane. J. Biol. Chem. 278, 12367-12373.

-       Kweon, D.-H., Kim, C. S., and Shin Y.-K. (2002) The membrane-dipped neuronal SNARE complex: A site-directed spin labeling electron paramagnetic resonance study. Biochemistry 41, 9264-9268.

-       Zhang, F., Chen, Y., Kweon, D.-H., Kim, C. S., and Shin, Y.K. (2002) The four-helix bundle of the neuronal target membrane SNARE complex is neither disordered in the middle nor uncoiled at the C-terminal region. J. Biol. Chem.  277, 24294-24298.

-       Kweon, D.-H., Chen, Y., Zhang, F., Poirier M., Kim, C. S., and Shin, Y.K. (2002) Probing domain swapping for the neuronal SNARE complex with electron paramagnetic resonance. Biochemistry 41, 5449-5452.

-       Kim, C. S, Kweon, D.-H., and Shin, Y.K. (2002) Membrane topologies of neuronal SNARE folding intermediates. Biochemistry 41, 10928-10933.

-       Kweon, D.-H., Lee, D.-H., Han, N. S., Rha, C.-S. and Seo. J.-H. (2002) Characterization of polycationic amino acids fusion systems for ion-exchange purification of cyclodextrin glycosyltransferase from recombinant Escherichia coli. Biotechnol. Prog. 18, 303-308.

-       Park, Y.-C., Kweon, D.-H., Lee, D.-H., and Seo, J.-H. (2001) Improved technologies to produce heterologous proteins in recombinant Escherichia coli. K. J. Biotechnol. Bioeng. 16: 1-10.

-       Jin, H.-H., Han, N. S., Kweon, D.-H., Park, Y.-C., and Seo, J.-H. (2001). Effects of environmental factors on in vivo folding of Bacillus macerans cyclodextrin glycosyltransferase in recombinant Escherichia coli. J. Microbiol. Biotechnol., 11, 92-96.

-       Kweon, D.-H., Han, N. S, Park, K.-M., and Seo, J.-H. (2001). Overproduction of Phytolacca insularis protein in batch and fed-batch culture of recombinant Escherichia coli. Process Biochemistry, 36, 537-542.

-       Kweon, D.-H., Lee, D.-H. and J.-H. Seo. (2001). Improved refolding strategy for active recombinant proteins. Biochemical Engineering. Protein folding/refolding. 15, 41-48.

-       Kim, C.-S., Han, N. S, Kweon, D.-H., and Seo, J.-H. (1999). Expression of Bacillus macerans cyclodextrin glucanotransferase in Bacillus subtilis, J .Microbiol. Biotechnol., 9, 230-233.

-       Lee, K.-G., Kim, M.-D., Kweon, D.-H., Han, N.-S., Park, K.-H. and Seo, J.-H.. (1999) Simultaneous expression of Bacillus licheniformis maltogenic amylase and thermostable amylase in recombinant Escherichia coli, Food Sci. Biotechnol. 8, 347-350.