HyunSook Lee, Ji Hye Lee, Min-Jin Kang, Md. Abdul Hannan, Il Soo Moon
언어
영어(ENG)
URL
https://www.earticle.net/Article/A192834
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원문정보
초록
영어
N-acetylglucosamine (GlcNAc) kinase (NAGK; EC 2.7.1.59) converts GlcNAc into GlcNAc-6-phosphate. Based on the proposed 3D structure, we produced 3 point mutant NAGKs that were expected to retain differential capacities for substrate binding and reaction velocity. The proteins were expressed in Escherichia coli, affinity-purified to homogeneity, and used for functional analysis. Among the mutants, conversion of Cys143, which does not make direct hydrogen bonds with GlcNAc to Ser (i.e., C143S) had the least affecton enzymatic activity. Conversion of Asn36, which plays a role in domain closure by making a hydrogen bond with GlcNAc to Ala (i.e., N36A) mildly reduced the enzyme activity. Conversion of Asp107, which makes hydrogen bonds with GlcNAc and acts as a proton acceptor to Ala (i.e., D107A), caused a total loss in the enzyme activity. The eGFP- or RFP (DsRed)-tagged mutant NAGKs increased the complexity of dendritic architecture when overexpressed in rat hippocampal neurons (DIV 5-9) with no statistical difference with wild-type NAGK. These results indicate that the upregulation of dendritic complex by NAGK is the enzyme’s non-canonical function.
저자
HyunSook Lee [ Department of Anatomy, Dongguk University College of Medicine, Gyeongju 780-714, Korea ]
Ji Hye Lee [ Department of Anatomy, Dongguk University College of Medicine, Gyeongju 780-714, Korea ]
Min-Jin Kang [ Department of Anatomy, Dongguk University College of Medicine, Gyeongju 780-714, Korea ]
Md. Abdul Hannan [ Department of Biotechnology, Pukyong National University, Namku, Busan 608-737, Korea ]
Il Soo Moon [ Department of Anatomy, Dongguk University College of Medicine, Gyeongju 780-714, Korea ]
본 학회는 화학, 생화학, 분자생물학, 미생물학, 식품공학, 의학, 약학, 유전공학 및 생물공학, 환경 및 기타 공업 등 전 분야의 탄수화물관련 이론과 기술을 연구 발전시키고 산학협동을 통해 이를 보급하여 국내 관련 산업의 발전 및 국민생활의 과학화에 기여하고자 하며, 이러한 목표와 비젼의 실현을 위해 회원들이 적극적인 참여와 활동을 전개하고자 한다.